Cartridge for automatic detection of analyte in body fluid sample and system comprising it

FIELD: medicine.

SUBSTANCE: group of inventions refers to laboratory diagnostics. A device for the automatic detection of an analyte in a body fluid sample comprises an array of addressed analysis units for carrying out a chemical reaction, which generates a distinguishable signal carrying the information concerning the presence or absence of the analyte; an array of addressed reagent units referring to the individual addressed reagent unit, corresponding to the individual addressed analysis unit, and wherein the individual reagent unit is configured to be calibrated by a reference signal in the respective individual analysis unit before the arrays are assembled on the device; the individual analysis unit comprises a quantitative analysis tip having an inner surface carrying the reagents fixed on the surface for the analyte detection. The group of inventions refers to an embodiment of the above device additionally comprising a sampling unit, as well as to sub-systems comprising the above devices, methods for analysing using the above devices, a method for assembling the above devices and a method for sampling plasma from the blood sample in these systems.

EFFECT: group of inventions provides the higher flexibility and reliability of the system, as well as prevents cross-contamination of the units of the device.

65 cl, 21 dwg, 3 tbl, 7 ex

Prerequisites to the creation of inventions

Detection of a large number of biomarkers of disease and the development and implementation of miniaturized medical systems has opened new ways to predict, diagnose and monitor the treatment of diseases in the place of the care. System located at the site of patient care, and helps to give the test results to medical staff, other health professionals and patients. Early diagnosis of the disease or the disease allows medical personnel to timely initiate or modify therapy.

Multiplex measurement of the biomarker provides additional information about the patient's condition. For example, when monitoring the effects of drugs three or more biomarkers can be measured in parallel. Typically, use titration microplates and other similar devices for a combined quantitative analyses based on separation. Titration microplate (for example, tetrazinni the microtiter plate with 384 wells) allows a large number of quantitative analyses in parallel.

In the device, located at the site of patient care (DFB device), the number held in parallel quantitative analysis is often limited by the device size and sample volume, parliamentarism. In many DFB devices to carry out more quantitative analysis is approximately from 2 to 10. It is desirable to have GROWN a device that allows the United quantitative analysis on a small sample volume.

The disadvantage of many ROS devices for the combined quantitative analysis is the high cost of manufacturing the components of the device. If the device is a disposable, high cost components can make use of the ROS device impractical. In addition, in the case of the United ROS device that contains all the necessary reagents within the device, if only one of these reagents will be unstable, it is necessary to discard the entire device, even if all other reagents may also be used.

When a consumer is interested in custom manufacturing GREW device with a specific set of analytes, then the manufacturers GREW up systems for the combined quantitative analysis are often faced with the necessity of mixing and matching (mix-and-match) quantitative assays and reagents devices. United GREW quantitative analysis, suitable for each user, can be very expensive, difficult to calibrate and difficult to maintain quality control.

It was proved that ROS methods are very anymopre monitoring of disease and therapy (for example, system for measuring blood glucose in the treatment of diabetes, measurement of prothrombin time in anticoagulant therapy using zoomarine). It can be assumed that due to the measurement of multiple markers can better control complex diseases (such as cancer) and therapy, such as therapy with multiple drugs.

The invention

Thus, considering the above there remains a need for alternative building GREW devices. Desirable design should be modular surface capture and incubation items for the quantitative analysis. Moreover, the modular surface capture and incubation items for the quantitative analysis necessary to build a GROWING consumable items, adapted for use in the methods of making and operational delivery of nodes. It would be desirable to create a custom GREW up device at a price acceptable to the customer and the manufacturer. The present invention allows to satisfy these needs, and provide appropriate benefits.

In accordance with the first aspect of the present invention, it is proposed cartridge for automatic detection of analyte in a sample of bodily fluid that contains: a matrix of addressable units of analysis that are configured for carrying out a chemical reaction, which is Aya gives a visible signal, carries information about the presence or absence of the analyte; a matrix of addressable blocks of reagents, which refer to individual adisoemarto block reagent matrix corresponding to the individual adisoemarto block analysis matrix units of analysis, and in which individual blocks of reagents configured for calibration reference signal corresponding to the individual unit of analysis, previously the Assembly of the matrices in (in) cartridge. The device may further comprise a block sampling, configured to receive a sample of bodily fluid.

In accordance with another aspect of the present invention, it is proposed cartridge for automatic detection of analyte in a sample of bodily fluid that contains: block sampling, configured to receive a sample of bodily fluid; matrix units of analysis that are configured to receive portions of the sample from unit sampling and for carrying out a chemical reaction, which gives a distinct signal that carries information about the presence of the analyte in the sample; the matrix blocks reagents containing reagents for chemical reactions; and the individual unit of analysis matrix units of analysis and the individual unit of the reagent matrix blocks reagents configured movable and having a fluid connection, so the reagents for conducting a chemical reaction in which can be entered in contact with the sample portion in the unit of analysis.

Individual block reagent may be configured to receive the rolling unit of analysis. In accordance with some variations, the individual unit of analysis contains (is a) a handpiece for quantitative analysis. In accordance with some variations, the individual unit of analysis is configured for carrying out immunological analysis.

The sample of bodily fluid may be the breakdown of the blood. In some cases, the unit of sampling is configured to receive a sample volume of bodily fluid comprising approximately 50, 20, 10, 5 or 3 μl or less. In particular, the unit of sampling is configured to receive a sample volume of bodily fluid, the equivalent of one drop of blood.

Described here, the device may have a preprocessing block configured to select portions of the sample of bodily fluid for chemical reactions to detect the analyte, and the preprocessing block can be configured for selection of plasma from samples of whole blood, entered in the unit of sampling.

In accordance with another aspect of the present invention, a system for automatic detection of analyte in a sample of bodily fluid that contains: a device suitable described here above; node detection, designed for detecting the supply distinct signal, carries information about the presence or absence of the analyte. The system may further comprise a programmable mechanical device configured to move the individual unit of analysis from the first location to the second location. In some cases, the system contains a device for transferring fluid. Device for transferring fluid may be a pipette, and can also be automated device. The system may also contain the node communication Protocol based on the analyte to be detected. In some cases, this system contains a heating unit configured to receive the individual unit of analysis, and also includes a magnetic block, for example, which can be used for separation of erythrocytes from blood samples.

In accordance with another aspect of the present invention, a system for automatic detection of multiple analytes in a sample of bodily fluid, which comprises: a fluid device, which contains: block sampling, configured to content (storage) of the sample of bodily fluid; matrix units of analysis, and the individual unit of analysis of this matrix blocks analysis is configured for carrying out a chemical reaction, which gives a signal that carries information about individuals who Inom the specified analyte many to be the detection of analytes; the matrix blocks, reagents, and individual block reagent specified matrix blocks of the reactants contains a reagent; a device for transferring fluid, which includes many heads, and individual head set head configured to enter into engagement with the individual unit of analysis, with the specified device for transferring fluid includes a programmable processing device, configured for direct transfer of the sample of bodily fluid from a block of sample and reagent from the individual unit of the reagent in the individual unit of analysis. In accordance with some options, the configuration processing device for direct transfer fluid allows you to set the degree of dilution of the sample of bodily fluid in the matrix blocks of the analysis, in order to receive signals carrying the information of the plurality of analytes to be detected within the detection range, so that the specified set of analytes can be detected using this system.

In some cases, the sample of bodily fluid contains at least two analyte present at concentrations that differ by at least 2, 5, 10, 15, 50 or 100 orders of magnitude. The degree of dilution of the sample of bodily fluid can give signals carrying the information of at least two of the analyte is within the detection range.

The proposed system may further comprise a detection unit configured to detect the intensity of the signal in the detection range. An exemplary detection device is a photomultiplier tube, the detection range is approximately from 20 to 10 million counts per second).

In accordance with some options, individual cylinder device to transfer fluid configured for coupling with the individual unit of analysis. The individual unit of analysis can provide location response immunological analysis. In some cases, the individual unit of analysis is the tip of the pipette. Device for transferring fluid may be a pipette, such as a pipette with the displacement of air. Device for transferring fluid may also contain a motor, connected to the programmable processing device, and the specified engine can move a specified set of heads on the basis of the record from the specified programmable processing device.

In accordance with another aspect of the present invention, a system for automatic detection of multiple analytes in a portion of the plasma sample of whole blood, which comprises: a device configured to automatically receive and about the abode samples of whole blood, to receive a portion of the blood plasma, with which the device generates a visible signal that carries information about the presence or absence of interest of the analyte; and node discovery, designed to detect visible signal, which carries information about the presence or absence of the analyte.

In accordance with another aspect of the present invention proposes a method of detecting analyte in a sample of bodily fluid, which includes the following: submission of blood samples in the above device; creating conditions for response specified sample within at least one unit of analysis; and detecting a specified audible signal generated by the specified analyte collected in said sample of bodily fluid. The breakdown of the bodily fluid may be a blood sample, and the method may include sampling (extraction) of plasma from blood samples.

In accordance with another aspect of the present invention proposes a method of Assembly at the request of the cartridge for automatic detection of analyte in a sample of bodily fluid, the device includes a housing, and the housing includes: a matrix of addressable units of analysis, in which the individual unit of analysis matrix configured for carrying out a chemical reaction, which gives discern the initial signal, carries information about the presence or absence of the analyte; a matrix of addressable blocks of reagents, which apply to the individual unit of the reagent matrix corresponding to the individual unit of analysis matrix, and the method includes the following operations: (i) placement in the body, in accordance with the subject to detect the analyte, matrix addressable units of analysis, and the individual unit of analysis matrix configured for carrying out a chemical reaction, which allows to detect interest of the analyte, the ordered specified by the end user; (ii) placement in the body, in accordance with the subject to detect the analyte, the matrix blocks, reagents, moreover, the individual unit of the reagent matrix corresponds to the individual unit of analysis; and (iii) the consolidation of the matrices (i) and (ii) inside the unit. The method may include the choice should be detected analyte. In accordance with some variations, the method involves sealing the cartridge. In one embodiment, the method involves marking cartridge readable label, which is subject to detection of the analyte, for example, in the form of a barcode or RFDD.

In accordance with another aspect of the present invention, it is proposed a method of automatic detection of multiple analytes in a sample is a bodily fluid, which includes the following: introduction of the sample of bodily fluid in the fluid device, and a fluid device includes: a unit of sampling, configured to content (storage) of the sample of bodily fluid; matrix units of analysis, and the individual unit of analysis of this matrix blocks analysis is configured for carrying out a chemical reaction, which gives a signal that carries information about the individual analyte specified set of detectable analytes; matrix blocks, reagents, and individual block reagent specified matrix blocks contains reagents reagent; connecting the individual unit of analysis using a device for transferring fluid; the transfer of the sample of bodily fluid from the block sampling in the individual unit of analysis using the device to transfer fluid; and transferring reagent from the individual unit of the reagent in the individual unit of analysis, in order to carry out the reaction of the reagent with a sample of bodily fluid and to receive a signal carrying information about the individual analyte many to be the detection of analytes.

In one embodiment, the device for transferring fluid contains many heads, with individual head set head configured to enter into engagement with the individual unit of analysis and the specified device for transferring fluid includes a programmable processing device, configured for direct transfer of the sample of bodily fluid from (from) block of sample and reagent from (from) individual unit of the reagent in the individual unit of analysis. The method may further include commands for programmable processing device, and specified commands can control the transfer operation of the sample of bodily fluid in the individual unit of analysis.

In one embodiment, the transfer operation of the sample of bodily fluid includes a change in the degree of dilution of the sample of bodily fluid in the individual unit of analysis, to obtain a signal that carries information about the individual analyte many to be the detection of an analyte within the detection range. The sample of bodily fluid can contain at least two individual analyte that is present at concentrations that differ by at least 2, 5, 10, 15, 50 or 100 orders of magnitude. In some cases, the degree of dilution of the sample of bodily fluid allows you to receive signals carrying the information of at least two individual analytes within the detection range. In one embodiment, the detection range is approximately from 1000 to 1 million samples per second, when using a photomultiplier.

In one embodiment, the reagent in the individual unit of the reagent is from the battle of the substrate of the enzyme for immunological analysis, and the method may further include repeating the operations of transferring reagent from the individual unit of the reagent, after completion of the reaction, to obtain a signal carrying information about the individual analyte many to be the detection of analytes, through which flows a second response to receiving the second signal, which carries information about the individual analyte. Signal intensity and the second intensity of the second signal carrying information about the individual analyte, average to get the final intensity of the signal carrying information on the individual analyte.

In accordance with another aspect of the present invention, it is proposed a method of measuring the volume of liquid samples (biological fluid), which includes the following operations: dealing with a known quantity of a control analyte in said sample with a reagent to obtain a visible signal that carries information about the number of control analyte; and comparing the specified visible signal with the expected distinguishable signal and the expected signal carries information about the expected volume of the sample, but such a comparison allows to measure the indicated amount of the indicated samples. In some cases, the control analyte is normally not present in the specified liquid sample in a detectable quantity. the manual may provide verification of the specified volume of the liquid sample, when the measured volume of sample is approximately less than 50% of the expected volume of the sample. In one embodiment, the method further includes: reacting with the reagent sample of bodily fluid containing the specified analyte to obtain a visible signal that carries information about a given analyte; and measuring the amount of a given analyte in a sample of bodily fluid using the specified intensity audible signal carrying information about a given analyte, and measuring the specified amount of the specified liquid samples. The liquid sample and the sample of bodily fluid can be the same sample, and the control analyte does not react with a given analyte in a sample of bodily fluid. In some cases, the liquid sample and the sample of bodily fluid represent different samples. The control analyte may be, for example, fluorescein-labeled (labeled with fluorescein) albumin, fluorescein-labeled IgG, anti-fluorescein, anti-digoxigenin, digoxigenin-labeled albumin, digoxigenin-labeled IgG, biotinylated proteins and IgG is not human.

In accordance with another aspect of the present invention, it is proposed a method of selection of plasma from blood samples, which includes the following operations: mixing of blood samples in the presence of magnetized particles in the unit of sampling, and nominitive the s particles contain surface of the capture antibody to bind Neoplatonic portions of the blood sample; and the imposition of the magnetic field over the area of collection of blood plasma for mixing blood samples, to obtain a suspension Neoplatonic portions of the blood sample on top of the collection zone of the plasma. In some cases, the unit of sampling is a capillary tube. The volume of blood samples may be approximately less than 20 μl, and the blood plasma is taken in the amount of approximately less than 10 μl. In some cases, the blood sample is not diluted. In some cases, the mixing occurs in the presence of antibody, unbound with a hard surface. Mixing may be mixing due to the action of the syringe.

In accordance with another aspect of the present invention, it is proposed a method of using an automated immunoassay for the detection of analyte present in the portion of the plasma sample of whole blood, which includes the following: submission of samples of whole blood in a device that is configured for automatic reception and processing of the sample of whole blood to obtain a portion of the blood plasma, giving a visible signal that carries information about the presence or absence of interest of the analyte; detecting a specified signal, which carries information about the presence or absence of analyte in said sample of bodily fluid; and sending results of operations the (b) end user. Immunological analysis can be analysis by ELISA. In some cases, the results are passed via wireless (radio).

In accordance with some of the options described here above process is carried out as described here above system.

All publications and patent applications mentioned in the description of the present invention, are included in this description by reference to the extent which corresponds to the volume of each individual publication or patent application.

These and other novel features and advantages of the invention will be more apparent from the subsequent detailed description, given with reference to the accompanying drawings.

Brief description of drawings

In Fig.1 shows an exemplary device in accordance with the present invention, which contains units of analysis the unit of reagents, and other modular components of the device.

In Fig.2 shows a cut from two sides of the exemplary device shown in Fig.1, which contains a cavity in the device, the shape of which allows you to enter the unit of analysis unit of the reagent and the tip of the sampling.

In Fig.3A shows the approximate analysis block, which contains a small tip or tube.

In Fig.3B shows the approximate tip for sampling.

In Fig.4A and 4B shows two example block the reagent, which contains the cap.

In Fig.5 shows an example of a system that holds the device in accordance with the present invention and the device for transferring fluid.

In Fig.6 shows an exemplary system in accordance with the present invention, which contains a heating unit for adjusting the temperature and the detection device.

In Fig.7 shows an exemplary system in which the patient enters the blood in the device, then the device is inserted into the reader.

In Fig.8 shows a block diagram of a system for evaluating a medical condition of the patient.

In Fig.9A-9F shows an example of a method of separating blood plasma, in which a sample of whole blood is drawn into the tip to the sample and the magnetic reagent is mixed with a sample and form a suspension, and then apply a magnetic field to the sample mixture of whole blood and the magnetic reagent. Then separated from the blood plasma is distributed in the wells of the device.

In Fig.10 shows an exemplary method of performing control of quantitative analysis, which use known number of control analyte.

In Fig.11 shows a thin film, such as film contamination inside the tip, when push one liquid and suck other liquid.

In Fig.12 shows a calibration curve correlating the unit of analysis and unit reagent is for the quantitative analysis for VEGFR2.

In Fig.13 shows a calibration curve correlating the results of the unit of analysis and unit of the reagent for quantitative analysis for P1GF in the system, when the measurement is performed using a luminometer.

In Fig.14 shows the concentration of CRP in the function signal of quantitative analysis (number of photons), and the data adjusted five-membered polynomial function to generate a calibration function.

In Fig.15 shows the obtained correspondence between the model and measurements described here, the parameters Smax, C0.5 and D.

In Fig.16 shows the data after dilution, used to achieve a final concentration in the tip for quantitative analysis.

In Fig.17 shows the normalized response of the quantitative analysis (In/Vth) as a function of the logarithm of the normalized concentration (C/C0.5), for the following relative dilutions: 1:1 (solid line), 5:1 (dashed line) and 25:1 (dotted line).

In Fig.18 and 19 shows an example similar to the one shown in Fig.17, when other normalized concentrations.

In Fig.20 shows the response of the quantitative analysis for the control analyte, after carrying out the following operations: delete antibody detection, flushing analysis and the substrate during reading using spectrofluorometry within 0.5 sec.

In Fig.21 shows achiev taty analysis, obtained by counting photons within approximately 10 seconds in the proposed system.

Detailed description of the invention

Described here are the options and aspects of the present invention relate to devices, systems and methods for automatic detection of analyte in a sample of bodily fluid. The present invention allows the detection and/or determination of the number of analytes that are associated with specific biological processes, physiological conditions, disorders (disorders) disorders or stages, or with the effects of biological or therapeutic agents. As described here, and the examples of the present invention are not intended to limit the scope of patent claims of the present invention.

Device

In accordance with the first aspect of the present invention, a device for automatic detection of analyte in a sample of bodily fluid, which contains a matrix of addressable units of analysis that are configured for carrying out a chemical reaction, which gives a distinct signal that carries information about the presence or absence of the analyte and the matrix addressable blocks of reagents, which refer to individual adisoemarto block reagent matrix corresponding to the one or more addressable units of analysis indicated in the data device, so that individual blocks of reagents can be calibrated by reference signal of the corresponding block (block) analysis of previously build matrices on (in) the device.

In accordance with another aspect of the present invention, a device for automatic detection of analyte in a sample of bodily fluid, which contains a matrix units of analysis that are configured for carrying out a chemical reaction, which gives a distinct signal that carries information about the presence of the analyte and the matrix blocks reagents containing reagents for carrying out a chemical reaction, and at least one of the blocks of the analysis and at least one of the blocks of reagents configured to move relative to each other within the device, so that the reagents for carrying out chemical reactions automatically entered into contact with a sample of bodily fluid in the unit of analysis.

In one embodiment, the device in accordance with the present invention, reference to the matrix units of analysis or blocks of reagents produced in accordance with the chemical reaction carried out using the configured unit of analysis. In another embodiment, at least one of the blocks of the analysis and at least one of the blocks of reagents configured to move relative to each other within the device, so that the reagents on what I'm carrying out a chemical reaction is entered automatically into contact with a sample of bodily fluid in the unit of analysis.

In one embodiment, the device in accordance with the present invention is self-contained and contains all the reagents, including reagents in liquid and solid phases, necessary for carrying out a variety of quantitative analyses in parallel. Optionally, the device may be configured for holding at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30,40, 50, 100, 200, 500, 1000 or more tests. Optionally, one or more control quantitative analyses can also be performed in the device in parallel.

Quantitative analysis can be quantitative immunological tests that can be conducted within a short period of time. Another type of quantitative analysis that can be performed in the device in accordance with the present invention, includes (but without limitation) measurement sequences (sequences of nucleotides) of the nucleic acid and measuring metabolites such as cholesterol. In accordance with some options, quantitative analysis completed within the time not exceeding one hour, and mostly less than 30, 15, 10 or 5 minutes. In accordance with other options, quantitative analysis is carried out in a period of time less than 5 minutes. The duration of the quantitative analysis can be adjusted depending on the type kolichestvennoj the analysis, which should be in the device in accordance with the present invention. For example, if a higher sensitivity, quantitative analysis can be carried out in the course of time more than one hour or one day. In some examples, the quantitative analyses that require long periods of time, are more practical to implement in other GROWING applications, such as in the home than in clinical applications GREW.

Any bodily fluid, which presumably contains interest of the analyte, can be analyzed in the system or device in accordance with the present invention. Usually used bodily fluids include, but are not limited to, blood, serum, saliva, urine, gastric and digestion fluids, tears, feces, semen, vaginal fluid, interstitial fluid, obtained from tumor tissue, and cerebrospinal fluid.

The bodily fluid can be withdrawn from the patient and introduced into the device through a variety of means, including (but without limitation) using tubes, injections or pipette. Used here, the terms subject and patient are used interchangeably, which is used to denote a vertebrate animal, preferably a mammal, and it is preferable to refer to one person. Mammals include, but are not limited to mice or rats, monkeys, humans, animals, sports and Pets. In one embodiment, the Lancet pierces the skin and takes samples using, for example, gravity, capillary action, suction or vacuum. The Lancet may be part of a device, or part of a system, or a separate component. If necessary, the Lancet can be powered using a variety of mechanical, electrical, Electromechanical or any other known mechanism actuation, or any combination thereof. In another embodiment, there is no active mechanism is not required, as the patient can enter bodily fluid in the device, for example, when introducing a sample of saliva. Collected fluid can be placed in the block sampling within the device. In accordance with another variant, the device comprises at least one Mitroglou to puncture the skin.

The volume of bodily fluid, which is used in the device, typically is approximately less than 500 μl, typically approximately from 1 to 100 μl. Optionally, samples from 1 to 50 μl, 1 to 40 μl, 1 to 30 μl, 1 to 10 μl, or even from 1 to 3 ál can be used for detection of the analyte using the device.

In one embodiment, the volume of bodily fluid, used for discovery the analyte in the claimed devices or systems, represents one drop of liquid. For example, one drop of blood from a pricked finger may form a sample of bodily fluid, the analysis of which is done using described here, the device, system or method.

The sample of bodily fluid can be withdrawn from the subject and introduced into the device in accordance with the present invention as described hereinafter in more detail.

In one embodiment, the matrix units of analysis and units of reagents configured in the form of a set of components obtained by mixing and fit (mix-and-match). The units of analysis can have at least one gripping surface, allowing it to react with the analyte from the sample of bodily fluid. The unit of analysis may be tubular tip with a gripping surface on the inside of the tip. Hereinafter described in more detail examples of the tips in accordance with the present invention. Block reagent typically stores a liquid or solid reagents needed to conduct the quantitative analysis, which allows to detect this analyte. Each individual block of the quantitative analysis and the unit of the reagent can be configured for independent operation of the quantitative analysis. For Assembly of the unit blocks are collected using the online hosts supply for use in the United cartridges.

Mouthbut made separate components of a liquid, and in the solid phase, after which examine their characteristics and sent to storage. In one embodiment, the assembling is carried out on request at the place of manufacture. The device may be modular and may include components such as a housing that is common to all quantitative assays, units of analysis, such as lugs and blocks reagents, such as different fragile or operating using the tool (instrument operable containers that capsulebuy containing liquid reagents. In some cases, the assembled device is then subjected to tests to verify the calibration of the dependence of the system response from the levels of known analyte). Device (blocks) analysis can be collected on request from the library of pre-built and calibrated items. In accordance with some variations, the fluid passages within the device can be simple and eliminates any possibility of the capture of the bubbles, as well as providing effective rinsing of the excess of labeled reagents when conducting quantitative analyses with excess reagents, such as ELISA assays.

The housing for the device in accordance with the present invention can be made of polystyrene or other moldable or subjected to machining plastic, and may have a set of conventions is to accommodate units of analysis and units of reagents. In one embodiment, the housing has means for draining (blotting) of the lugs or blocks analysis to remove the excess liquid. Means for draining may be a porous membrane, such as cellulose acetate, or a piece of absorbent material such as filter paper.

In accordance with some variations, at least one of the components of the device may be made of a polymeric material. As non-restrictive examples of polymeric materials can lead to polystyrene, polycarbonate, polypropylene, polydimethylsiloxane (PDMS), polyurethane, polyvinylchloride (PVC), polysulfone, polymethylmethacrylate (emission spectra obtained for pure), a copolymer of Acrylonitrile, butadiene and styrene (ABS) and glass.

Device or subcomponents of the device can be manufactured using various methods, including (but without limitation) by stamping, injection molding, extrusion (embossing), casting, blow molding, machining, welding, ultrasonic welding and thermal welding. In one embodiment, the device is manufactured using injection molding, thermal welding and ultrasonic welding. The subcomponents of the device can be attached to each other by thermal welding, ultrasonic welding, friction fit (fits under pressure), using adhesives Il is, in the case of some substrates, for example, of glass, or of semi-rigid and flexible polymer substrates, due to the natural adhesion between the two components.

An exemplary device, such as described here, is shown in Fig.1. The device 100 also sometimes called here the cartridge 100. The device 100 includes a housing 130 with the locations for placement of blocks 121 and analysis units 103, 122, 124, 125 reagents. In the exemplary embodiment shown in Fig.1, the blocks 121 analysis is Central to a number of housing 130 of the device 100. Blocks 121 analysis may optionally contain at least one block 126 calibration. In accordance with one example, the blocks 121 analysis similar to the pipette tip and can be called lugs 121 for quantitative analysis, and blocks 126 calibration can be called lugs 126 calibration, but it should be borne in mind that the blocks 121 analysis can be of any shape and size, allowing you to enter them in as described here, the device 100. Blocks 121 analysis and blocks 126 calibration are approximate blocks 121 analysis and is described in more detail below. Blocks 121 of the analysis, shown in Fig.1, may have a gripping surface and allow, for example, to carry out a chemical reaction, for example, necessary for the implementation of the quantitative analysis of nucleic acid and immunological analysis of the century. Blocks 121 analysis can be collected in the housing in accordance with the instructions of the user or in accordance with the quantitative analyses that the user wishes to perform on the test.

As shown in Fig.1, the body 100 of the device may have an array 110 of sampling, configured to content of the sample. Sample such as a blood sample, can be placed in block 110 sampling. The tip 111 of sampling (for example, the tip of the pipette, which is connected with a device for transferring fluid, as described in more detail below) may take other part of the body 130. When carrying out the quantitative analysis, the tip 111 of sampling can distribute the sample into blocks pre-treatment reagents or blocks 103, 104, 105, 106, 107 pre-treatment, or in blocks 121 analysis. Approximate blocks 103, 104, 105, 106, 107 pre-treatment include (but without limitation): blocks 107 mixing, blocks 103, 104 dilution and, if the sample is a blood sample, blocks 105, 106 selection of blood plasma. Blocks 103, 104, 105, 106, 107 pre-treatment may be units of the same type or of different types. Experts can easily understand that other blocks 103, 104, 105, 106, 107 pre-processing necessary for carrying out chemical reactions, can also be built into the device 100. Blocks 103, 104, 105, 106, 107 can in order to win various amounts of reactants or diluents, necessary to perform the quantitative analysis in this cartridge 100.

Blocks 121 analysis can often be manufactured separately from the housing 130 and is then inserted into the housing 130 by means of the methods of capture and installation (pick-and-place). Blocks 121 analysis can be tightly inserted into the housing 130 or can be freely inserted into the housing 130. In accordance with some variations, the housing 130 is made so that it tightly holds the blocks 103, 122, 124, 125 reagents and/or blocks 121 on-site analysis, for example, during shipment or movement of the cartridge. Blocks 103, 122, 124, 125 reagents shown in Fig.1, contain conjugated reagent 122 (e.g., for use in immunological analysis), wash reagent 125 (for example, for washing the specified conjugate with the surface of the grip) and the substrate 124 (for example, the substrate of the enzyme). The following describes other options device 100 and components in accordance with one example in Fig.1. Blocks 103, 122, 124, 125 reagents can be manufactured and filled (reagents) separately from the housing 130 and is then accommodated in the housing 130. Due to this, the cartridge 100 can be made modular, which increases the flexibility of use of the cartridge 100 for various quantitative analyses. The reagents in block 103, 122, 124, 125 reagent can be selected in accordance with the ongoing quantitative analysis. Next, description is s in more detail an exemplary reagents and quantitative analyses.

A device such as shown in Fig.1, may also have other characteristics necessary for carrying out chemical reactions. For example, if the blocks 121 of the analysis are described here lugs 121 for quantitative analysis, then the device may contain a dewatering pad (touch-off pads 112 to remove excess sample or reagent from the tip 121 for quantitative analysis or tip 111 sampling after transfer fluid, for example, using the system described here. The housing 130 may also be blocks or areas 101, 102 within the device 100 to accommodate the used tip or block, for example, to avoid cross-contamination of the tip 111 of sampling or block 121 analysis. In Fig.1 shows a device 100 that contains the tip 111 of sampling, designed to move the sample between the blocks of the device 100. The device 100 shown in Fig.1, also includes a tip 113 pre-treatment, designed to move the sample, which was pre-treated in the same block device 100, in other blocks device 100 for carrying out chemical reactions. For example, the tip 111 of the sampling can be used for sampling blood from the block 110 sampling and to move blood samples in blocks 103, 104, 105, 106, 107 pre-treatment is otki, as mentioned here above. Red blood cells may be removed from blood samples in blocks 103, 104, 105, 106, 107 pre-treatment and the tip 113 pre-processing can then be used to collect blood plasma of the blocks 103, 104, 105, 106, 107 pre-treatment and transfer plasma into another block 103, 104, 105, 106, 107 pre-treatment (for example, in block dilution) and/or at least one unit 121 analysis. In one embodiment, the tip 111 sampling is a block 110 sampling. In another embodiment, block 110 sampling is similar to the hole and can be configured to store samples received by the user.

Blocks 121 and analysis units 103, 122, 124, 125 reagents shown in Fig.1, may have an available address to specify the location of the blocks on the cartridge 100. For example, one column of the cartridge 100, as shown in Fig.1, may contain block 121 of analysis, intended for the quantitative analysis, configured to detect C-reactive protein, and the other columns can contain the corresponding blocks 103, 122, 124, 125 reagents for quantitative analysis in the above mentioned column, and these blocks are made with recourse to each other. For example, addresses can be entered using a computer system and can granites is in it, as the cartridge 100 may have an appropriate label, such as a bar code. After scanning the barcode of the cartridge 100, the computer can send the addresses of blocks declared in the system to move the fluid and to conduct the reaction in accordance with the addresses entered in the computer. Addresses can be part of the Protocol that controls the operation of the system. Addresses can be in any configuration and can be changed if you want to change the Protocol for quantitative analysis or operation the use of a cartridge that usually is not possible in previously known DFB devices. In accordance with some options, the units in the housing 130 to form the matrix 6 to 8 blocks, as shown in Fig.1. The format of the location of the blocks can be any, for example, it can be a rectangular matrix or a random layout. The cartridge 100 can have any number of blocks, for example approximately from 1 to 500. According to some variants, the cartridge 100 is 5-100 units. In the example shown in Fig.1, the cartridge 100 has 48 blocks.

In Fig.2A and 2B shows a cut from two sides of the exemplary device shown in Fig.1. In the housing 220 of the device may be formed cavity for placement of blocks 201 analysis (for example, tips for quantitative analysis) in a vertical orientation (horizontal case) base, education is built up in the device 200. As shown in Fig.2, can also be formed cavity for placement of the block 210, 212 reagent or block sampling or tip 202. The housing 220 may be made so as to accurately capture blocks and firmly to keep them. These characteristics also apply to the mechanism for moving tips, which produces the capture and installation (drop-off, reset) tip. In another embodiment, the block sampling contains bend or break-away element, which serves to protect a small team of the tube during shipment and to hold the plunger inside the capillary. Also in Fig.2A shows two exemplary variant of the claimed blocks 210, 212 reagents. The base 220 may be configured to collect liquid waste, for example washing fluid after use, when they flow into the base through the opening in the housing 220. The housing 220 may have an absorbent pad for collecting liquid waste. The units of analysis blocks 201 and 202 of the sampling can be passed through the body cavity 220 of the device 200 and may extend beyond the inner support structure. Blocks 210, 212 reagents tightly fixed in the housing, as shown in Fig.2, and not protruding beyond the inner support structure. The housing 220 and the zones in which established and held blocks 201 analysis and the blocks 210, 212 reagents can have a different configuration is s.

In accordance with some options, each lug serves to conduct the only quantitative analysis and can be paired with the appropriate reagent, or may comply with the appropriate reagent, which is required for carrying out the specified quantitative analysis. Some tips are designed for the control units of analysis and are a known quantity of the analyte on the surface of the grip caused in the manufacturing process of the lugs or quantitative analysis. In the case of a control unit of analysis, this unit is configured to carry out the monitoring of the quantitative analysis carried out for comparison. The control unit of analysis may be, for example, the engagement surface and the analyte in the liquid or solid state.

In many cases, the device contains all reagents and fluids that are required for quantitative analysis. For example, to conduct luminogenic (luminogenic) quantitative analysis of ELISA reagents within the device may contain a diluent sample, the means of detection conjugate (for example, three labeled enzyme antibodies), wash solution and the substrate of the enzyme. If necessary, can be added reagents.

According to some variants, the reagents can be introduced into the device d is I pre-treatment samples. As examples of reagents for pre-treatment can lead to (but without limitation) reagents lysis of cells, reagents for release of analytes from the matrix materials in the sample, enzymes and detergents. Reagents for pre-treatment can also be added to the diluent, which is contained within the device.

Individual block reagent may be configured to receive the rolling unit of analysis. In accordance with some variations, the individual unit of analysis is a hollow cylindrical member with an open end, which contains the engagement surface and the reaction cuvette. A cylindrical unit of analysis can also be called a tip for quantitative analysis. In accordance with some variations, the individual unit of analysis is configured for carrying out immunological analysis. Block 301 analysis, which contains a small tip or tube shown in Fig.3A. In some cases, the tip 301 is configured so that it has an internal cylindrical surface 311 capture and boss 321 made with the possibility of entrance into engagement with the housing of the device. In some cases, the boss 321 and the tip 301 is configured with the ability to enter into engagement with the transfer mechanism of the handpiece 301, such as described here is a system or for example, a device for transferring fluid. Tip 301 for the quantitative analysis shown in Fig.3A may have a hole 331 at the lower end. This hole 331 may be used to transfer fluids or reagents in the unit of analysis and from him. In one embodiment described here, the block 301 of the analysis is similar to the tip of the pipette, however, the block 301 analysis further comprises surface 311 capture, configured to detect the analyte in the sample.

Tip 301 may be manufactured using a molding process. In one embodiment, the tip 301 is made of transparent polystyrene, which is usually used in quantitative chemiluminescent assays. As shown in Fig.3A, an exemplary handpiece 301 contains a boss (shown as a wider upper part of the tip 301), which is made with the possibility of entrance into engagement with the body and may, for example, to engage with the conical elements of the device for transferring fluid and/or pipettes, with the formation of the hermetic seal. In addition, as shown in Fig.3A, an exemplary handpiece 301 has a small cylindrical part. In many cases, the engagement surface is inside this small cylindrical part, however, the engagement surface may also be in any other place within nakonec the ICA 301 or outside from the tip 301. The handpiece 301 may have any geometric shape, including (but not limited to, tubular, cubic or pyramidal. Based on chemiluminescence and fluorescence quantitative analyses tip 301 is a convenient tool for product introduction quantitative analysis in the optical path of the device.

In Fig.3C shows an exemplary block 302 sampling, which contains the tip 302 of sampling. It is shown in Fig.3B, the tip 302 of sampling may also be performed separately from block 302 sampling and can be used to transfer samples from block sampling in other blocks of the claimed device. It is shown in Fig.3B, the tip 302 of the sample contains a boss 322 to connect the handpiece 302 with the case of the claimed device and with a device for transferring fluid. Tip 302 sampling also has a hole 332, allowing for the introduction of fluids or samples in the tip of the sampling and remove them from it. In accordance with some variations, the tip 302 of the sample has the same form as the tip 301 for quantitative analysis. In other embodiments (such as shown in Fig.3A and 3B), the tip 302 of the sample has a different shape than the tip 301 for quantitative analysis.

In one embodiment, one of the functions of the tip is introduction the s contact of sample and liquid reagents to the surface of the capture unit of analysis. Moving can be done through a variety of means, including (but without limitation) using capillary action and suction controlled injection pump. The small size of the tips allows you to quickly adjust the temperature necessary for carrying out chemical reactions. The heat transfer and/or temperature maintenance can be carried out simply by placing the tip in the block temperature adjustment.

In accordance with some variations, the tip may contain approximately 1 to 40 μl of fluid. In accordance with other options, the tip may contain approximately 5 to 25 μl of fluid. In one embodiment, the tip contains 20 μl of fluid. In some cases, the tip may contain 1 μl of fluid or less. In accordance with other options, the tip can contain up to 100 μl of fluid.

If necessary, the tip of the handpiece can be dried by contact with an absorbent material (for example, integrated in a disposable cartridge), previously the introduction of the next component in the quantitative analysis, to avoid contamination of a small amount of sample and/or reagent. Due to the physical forces any liquid Tenuta in this tip, may be held in any desired place, with minimal risk of leakage of the liquid, even when UD is the neigh of a handpiece in a vertical position.

The unit of analysis (e.g., tip for quantitative analysis) can be covered before it is used reagents capture, using the same tools as in the quantitative analysis (for example, by means of an adjustable capillary or mechanical suction).

The engagement surface (also referred to here as the place of the reaction) can be formed due to binding of antibodies or other capture reagents associated covalently or by absorbing unit of analysis. The surface can then be drained and can be maintained in a dry state until use in quantitative analysis. In one embodiment, there is a place of reaction for each of the measured analyte.

In one embodiment, the unit of analysis can be displaced for insertion in fluid communication with the reagent and/or with block sampling, so that the reagent or the sample can interact with the site of reaction, where the associated samples can detect interest of the analyte in the sample of bodily fluid. Place the reaction then create a signal that carries information about the presence or concentration of interest of the analyte, which can then be detected by these detection devices.

In accordance with some options, the location and configuration of m is a hundred reactions are important elements in the device for quantitative analysis. Most, if not all, disposable device for immunological analysis performed so that the engagement surface is an integral part of the device.

In one embodiment, molded plastic unit of analysis may be purchased or can be made using injection molding, with the exact size and shape. For example, he may have a diameter of 0.05 to 3 mm and a length of from 3 to 30 mm Blocks can be coated with capture reagents using a method similar to that used for coating titration microplate, but with the advantage that it can be done processing in bulk, in a large tank by introducing a covering of reagents and processing using baskets, holders, etc. to extract the details and, if necessary, to rinse.

The unit of analysis forms a rigid support, which may be fixed reagent. The unit of analysis also has the appropriate characteristics of the interaction with light. For example, the unit of analysis can be made of such material, as a special (functionalized glass, Si, Ge, GaAs, GaP, SiO₂SiN₄modified silicon, or any material from a wide variety of gels or polymers such as (poly)tetrafluoroethylene, (poly)vinylidenefluoride, polystyrene, polycarbonate, polypropylene, emission spectra obtained for pure, ABS, or combinations thereof. One in which the Rianta, the unit of analysis contains polystyrene. In accordance with the present invention can be used, and other suitable materials. Transparent place of the reaction is preferred. In addition, in the case when there is an optically transmissive window that allows light to reach the photodetector, then the surface can mainly be opaque and/or predominantly light-scattering.

The reagent fixed on the surface of the grip may be any reagent useful for the detection of interest of the analyte in the sample of bodily fluid. Such reagents include (but without limitation) of the sample nucleic acids, antibodies, cell membrane receptors, monoclonal antibodies and antisera reactive relative to the specific analyte. Can be used in a variety of commercially available reagents, such as containing Polikanov and monoclonal antibody reagents specifically designed for specific analytes.

Experts can easily understand that there are various ways of fixing (immobilization) of various reagents on a support, where the reaction takes place. Immobilization may be covalent or non-covalent, at the expense of the share of the linker or due to their binding to immobilized share. As non-restrictive examples of connecting the components of the clients for attaching nucleic acid or protein molecules, such as antibodies to a solid support can lead to, among other things, by streptavidin or avidin/Biotin, urethane communication, ether, amide, diolefine of communication due to the (N)-functional thiourea, communication through the functional maleimide, amine, disulfide, amide and gidrazonami communication. In addition, the share of solila can be attached to nucleic acid for direct attachment to the substrate, such as glass, using known methods. Surface immobilization can also be obtained using a halyard (tether) role-L-lysine, which provides connection charge-charge with the surface.

The units of analysis can be dried after the last injection (injection) surface capture. For example, drying can be performed by passive exposure to a dry atmosphere or through the use of a vacuum manifold, and/or by passing clean, dry air through the collector.

In many cases, the unit of analysis is designed so that it can be used in mass production processes with high performance. For example, the tips can be installed in large matrices for the mass of coating on the surface of the grip or on the tips. In another example, the tips can be placed on a belt conveyor or carouse and for batch processing. In another example, a large matrix of tips to simplify the processing may be connected to a vacuum manifold and/or under pressure header.

In one embodiment, the unit of analysis may be operatively associated with a device for transferring fluid. Device for transferring fluid can operate with automatic control, without human intervention. In the case of units of analysis that contain tips, control the installed height of the disposable tip is based on taper attachment with tension tip to the liquid distribution. Device for transferring fluid can enter into engagement with the tip. In some cases, the depth of immersion of the tip into a portable liquid must be known in order to minimize contact with the outside of the tip, which can be uncontrolled. To join or to bite the bullet tip with a device for transferring fluid at the base of the conical connector may be formed of a solid support, which is engaged with a nozzle of the dispenser. Airtight seal may be formed through the sealing ring, which is located at half the height of the cone or on the flat base of the nozzle. Due to the separation of the functions of the sealing tip from a controlled height Nakonechny the ka, you can separately make their adjustment. Modular claimed device and the device for transferring fluid can produce many quantitative analyses in parallel.

Blocks of reagents device can store the reagents that are required for the chemical reaction to find this of interest of the analyte. Liquid reagents can be bottled in small capsules that can be made of various materials, including (but without limitation) of plastics, such as polystyrene, polyethylene or polypropylene. In accordance with some options, blocks of reagents contain cylindrical glasses. Two examples of block 401, 402 reagent, which contains the glasses shown in Fig.4A and 4C. If necessary, blocks 401, 402 can fit to enter into the cavity in the device. Blocks 401, 402 can be sealed on the outside surface to prevent splashing of reagents 411, 412 within the device. According to some variants, the glass sealing using aluminized plastic by thermal welding. The block may be of any shape, allowing you to store the reagent. For example, in Fig.4A shows a block reagent 401 cylindrical form, which can store liquid reagent 411. It is shown in Fig.4B block reagent 402 other forms also allow you to plug the et to store liquid reagent 412. Both exemplary block 401, 402 reagents, if necessary, may have slight modifications in the vicinity of the upper surface, allowing tightly to enter the blocks 401, 402 in the device, as mentioned here above.

In accordance with the variations in implementation of the present invention, blocks of reagents are modular. Block reagent is designed so that it can be used in mass production processes with high performance. For example, multiple blocks of reagents can be filled and sealed simultaneously in a mass process. Blocks of reagents can be filled in accordance with the type of quantitative analysis or quantitative analyses carried out using the device. For example, if one user wants to spend other quantitative analyses than another user, then the blocks of reagents can be manufactured in accordance with the preferences of each user, without the need of making another whole device. In another example, blocks of reagents can be placed on a belt conveyor or carousel for serial processing.

In another embodiment, the blocks of the reagents introduced directly into the cavity in the device. In this embodiment, the seal may be made in the areas of housing, surrounding blocks.

Reagents in accordance with aseason invention include (but without limitation) wash buffer solutions, the substrates of enzymes, buffer solutions for cultivation, conjugates labeled with enzyme conjugates, amplifiers DNA, sample diluents, wash solutions, reagents for pre-treatment of samples, including additives, such as detergents, polymers, chelating additives, albumin binding reagents, enzyme inhibitors, enzymes, anticoagulants, agglutinating agents of red blood cells, antibodies, or other materials necessary to conduct quantitative analysis on the device. Labeled enzyme conjugate can be policlonal antibody or monoclonal antibody labeled with an enzyme that allows you to create a distinguishable signal by reaction with a suitable substrate. As examples of such enzymes can lead to (but not limited to, alkaline phosphatase and horseradish peroxidase. According to some variants, the reagents are reagents for immunological analysis. Typically, reagents, particularly those that are relatively unstable when mixed with liquid, is stored separately in the specified area (for example, block reagent) inside the unit.

In accordance with some options, block reagent contains approximately from 5 µl to 1 ml of liquid. In accordance with some options, block reagent may contain approximately 20-200 ál of liquids is I. In accordance with another variant, the block reagent contains 100 μl of fluid. In one embodiment, the reagent contains about 40 μl of fluid. The volume of liquid in the block reagent may vary depending on the type conducted quantitative analysis or from the amount of available sample of bodily fluid. In one embodiment, the volumes of reagents are not specified, but must exceed the known minimum. According to some variants, the reagents are stored in dry form and dissolve before conducting quantitative analysis on the device.

In one embodiment, the blocks of reagents can be filled using a siphon, funnel, pipette, syringe, needle, or combinations thereof. Blocks of reagents can be filled using the channel fill and channel vacuum suction. Blocks of reagents can be completed individually or in a mass production process.

In one embodiment, the individual unit of the reagent contains a different reagent as a means of isolation reagents from each other. Blocks reagents may also contain a wash solution or the substrate. In addition, blocks of reagents may contain luminogenic substrate. In another embodiment, the block reagent contains a lot of chemicals.

In some cases, the device setting provides preliminary calibration b the shackles of analysis and units of reagents, previously, the Assembly of disposable components of this device.

System

In accordance with one aspect, the system in accordance with the present invention provides a device containing units of analysis and units of reagents, which contain the reagents (reagents both in liquid and solid phase). In accordance with some options, at least all the device, the unit of analysis unit of the reagent or combination of them are disposable. In the system in accordance with the present invention, detection of the analyte using a device made by the meter. In most cases, the meter, the device and method allow you to get an automated detection system. Automated detection system can be automated on the basis of the specified Protocol or Protocol entered into the system by the user.

In accordance with one aspect, a system for automatic detection of analyte in a sample of bodily fluid contains a device or Chuck, as well as the host of discovery or detection device, for detecting the visible signal, which carries information about the presence or absence of the analyte.

In one embodiment, the user enters the sample (e.g., measured or nezamechennoy the blood sample in the device and inserts the device into the meter. All subsequent operations is the fast automatic programmable using the meter (rigid installation), user, remote user or system, or modification of the meter is operating in accordance with the identifier (e.g. barcode or RFID device).

As examples of the various functions that can be implemented using the system in accordance with the present invention, one can cite (but without limitation) the dilution of the sample, removal of parts of the sample (e.g., erythrocytes), the test samples in the block of the analysis, the additive liquid reagents to the sample and unit of analysis, leaching reagents from the sample and unit of analysis, and storage of fluids during use and after use of the device. The reagents can be introduced inside the unit in the unit reagent or block reagent, which is injected into the device at the Assembly.

An automated system can detect a specific analyte in a biological sample (e.g. blood sample) by quantitative analysis with an associated enzyme immunosorbent assay (ELISA). The system is amenable to multiplexing and is particularly well suited for the detection of interest of the analyte present in a small sample volume of whole blood (for example, 20 μl or less). The system also allows you to detect analytes at various dilutions which have a unique samples that allows you to test different sensitivity to the same device. All reagents, supplies and waste can be stored in the system.

When using a sample from the subject injected into the assembled device, after which the device is introduced into the meter. In one embodiment, the meter can start processing samples through some combination of removal of red blood cells (in the case of blood samples), dilution of the sample and move the sample into the unit of analysis. In the variant with combined quantitative analyses using multiple units of analysis and the portion of the sample move in individual units of analysis in series or parallel. Can then be conducted quantitative analyses using a controlled sequence incubation and application of reagents on the surface of the grip.

An exemplary device for transferring fluid contains all the components required for the quantitative analysis and/or read the results of the quantitative analysis. As an example, components can lead to (but without limitation) pumps for suction precise amounts of fluid from the wells or block device, at least one cascade of forward movement, allowing to improve the accuracy of the navigation system, the device detecting the analyte in the EN bloc is Lisa and temperature controller, to regulate the temperature of the medium for incubation quantitative analyses. In one embodiment in accordance with the present invention, the meter regulates the temperature of the device. In accordance with another variant, the temperature is in the range of about 30-40 degrees Celsius. In accordance with some variations, the temperature control system provides active cooling. In some cases, the temperature range is approximately 0-100 degrees Celsius. For example, for quantitative analysis of nucleic acids, temperatures can reach 100 degrees Celsius. In one embodiment, the temperature range is approximately 15-50 degrees Celsius. The unit of the temperature control system may include a thermoelectric device such as a Peltier device.

Described herein cartridges, devices, and systems have many features that are missing in existing ROS systems or systems combined analysis. For example, many GREW cartridges use a closed fluid system, or a loop that allows efficient processing of small volumes of liquid. Described herein cartridges and fluid devices provide free flow of fluid between the blocks of the cartridge. For example, the reagent can be stored in the block (reagent), the sample in the loke sampling, the diluent is in the unit of a diluent, and the engagement surface may be in the unit of analysis, and, in one of the States of the cartridge, or one of the blocks has no fluid communication with other blocks. When using these devices to transfer fluid or system blocks in this state do not have fluid communication with each other. Blocks can be moved relative to each other in order to create a fluid connection between certain blocks of each other. For example, a device for transferring fluid may have a head that engages with the unit of analysis and introduces the unit of analysis in fluid communication with the reagent.

Described here are devices and systems effectively provide high performance and detection in real time of analytes present in a bodily fluid of the subject. The proposed detection methods can be used in a variety of circumstances, including for the identification and quantification of analytes that are associated with specific biological processes, physiological conditions, disorders (disorders) disorders or stages. As such, the proposed system have a wide range of use, for example, for drug screening, diagnosis, phylogenetic classification is AI, paternity and forensic identification, the beginning and repeat diseases, individual response to the treatment of different population groups and for monitoring therapy. Offered here is a device and system is also particularly useful for the assessment of preclinical and clinical stages of treatment to improve adherence to medical treatment of a patient, monitoring of erythrocytes associated with the designated drug for the development of individualized medicine, and allowing review of blood from the Central laboratory to the test at home or on the basis of the recipe, and monitoring of therapeutic agents after receiving permission for their use. Devices and systems allow you to create a flexible system for personalized medicine. When using the same system the device can be replaced together with the Protocol or commands for programmable processing device systems for the realization of a wide variety of quantitative analyses. The proposed systems and devices can be used in the laboratory, in the form of a table of automated meter is small in size.

In accordance with some options, the patient can have many devices used to detect different analytes. The patient may, for example, to use different f widnae devices on different days of the week. In accordance with some options, the program on an external device that associates the identifier with a Protocol may contain a process for comparing the current day from the day when it is necessary to use a fluid device, for example, on the basis of clinical examination. In another embodiment, the patient has the different blocks of the reactants and the units of analysis, which alternately can be entered into the device. In yet another variant, if the patient does not need a new device for each day of the test, the system can be programmed or reprogrammed by downloading a new command, for example, from an external device, such as a server. If, for example, two days of the week are not the same, the external device via radio sends a notification to the patient about how to choose the device and/or what to choose for the system. This is only one example, and in another case, a patient may receive a notification that the fluid device was not used at the right time of day.

For example, the cartridge, such as shown in Fig.1, can contain multiple units of analysis and units of reagents. The units of analysis may have a gripping surface in accordance with the subject to detect the analyte. The units of analysis can then be quickly (in just-in-time fashion) United with the rest of the disorder. In many previously known DFB devices gripping surface is integral with the device, and if the engagement surface formed incorrectly, the entire device is bad. When using the device described here can be made to order and control the surface quality of the capture and/or unit of analysis individually and independently of the blocks of the reagents and of the device.

Blocks reagents promptly can be filled with different reagents. This provides the flexibility of the device, manufactured to order. In addition, blocks of reagents can be filled with different volumes of reagents that do not affect the stability of the device or on a chemical reaction taking place inside the device. When connecting to described here, the system having a device for transferring fluid, the described devices and units provide flexibility in the implementation of methods and protocols conducted quantitative analyses. For example, a party similar devices containing the same reagents may be transferred to a pool of patients for clinical examination. After half of the clinical examination, the user finds that quantitative analysis can be optimized by changing the dilution and the amount of reagent introduced into the unit of analysis.After this quantitative analysis can be modified or optimized only due to the change command, supplied in a programmable processing device, which is part of a device for transferring fluid. For example, a consignment of ammunition in the pool of patients have an excess of diluent in the Chuck. Then the new Protocol requires 4 times thinner than the previous Protocol. Through the use of the proposed methods and systems, the Protocol can be modified in a Central server and sent to all systems for implementing the methods, devices, without the need to supply new device pool of patients. In other words, as described here GREW device and system provide much greater flexibility standard laboratory practice, when there are often excessive excess reagents and samples.

In some cases, when the blocks of the cartridge are separate devices and systems provide flexibility when designing systems described here. For example, the cartridge may be configured to conduct 8 quantitative analysis using matrix units of analysis and matrix blocks reagents. Due to the characteristics described here Chuck, the same case, or the case of the same design can be used for the manufacture of the cartridge, allowing up to 8 quantitative analysis other than in the previous bullet. Such flexibility is difficult to obtain in many well-known this is currently DFB devices due to the use of closed systems and fluid channels, so that these devices cannot be modular and easy Assembly.

Currently, there is a need to detect multiple analytes that are present in the range of greatly varying concentrations, for example, when one analyte is in the range of concentrations PG/ml, and the other analyte is in the range of concentrations µg/ml Described here, the system allows simultaneous quantitative analysis of analytes that are present in the same sample in the range of greatly varying concentrations. Another advantage associated with the ability to conduct quantitative analysis of analytes that are present in the range of greatly varying concentrations, is able to associate the relationship of the concentrations of these analytes with safety and efficiency purposes a variety of medicines to the patient. For example, unexpected drug interactions with one another is uncommon adverse reactions drug. Conducted in real-time, simultaneous measurement of different analytes allows you to avoid the potentially dangerous consequences of adverse drug interactions with one another.

The possibility of monitoring the rate of change of the analyte concentration and/or concentration is AI PD or RK markers within a period of time on a single patient, or trend analysis to the analyte concentration and/or concentration of PD or RK markers, when concentrations are the concentrations of drugs or their metabolites, can prevent a potentially dangerous situation. For example, if glucose is the interest of the analyte, the concentration of glucose in the sample at a given time and the rate of change of glucose concentration in a given period of time are very useful, for example, in the prediction and prevention of hypoglycemic events. This analysis of the trend of concentration is very useful when selecting the dosage of medicines. As for many drugs and their metabolites, it is often desirable to analyze the trend and the possibility of taking preventive measures.

Thus, the data obtained through the use of the proposed fluidic devices and systems may be used to perform a trend analysis of the concentration of analyte in a patient.

Very often an 8 quantitative analyses in the same cartridge may require different dilutions or pre-treatments. The range of dilution can vary between quantitative analyses. Many well-known ROS devices often have a limited range of dilution and therefore out a limited number of quantitative analyses, which potentially can be carried out in such a DFB device. However, as described here, the system and/or the cartridge have a wide range of dilutions due to the possibility of a serial dilution of the sample. Consequently, a large number of potential quantitative analyses can be performed on a single cartridge or set of cartridges, without requiring modification of the device detection or reading of the meter for quantitative analyses.

In accordance with one example, the proposed system is configured for a set (for example, five or more) defined quantitative analysis of the analyte. In order to bring the expected concentration of the analyte inside the detection range due to immunological analysis, commonly used in the field GREW, the sample must be diluted, for example, in the ratio 3:1, 8:1, 10:1, 100:1 and 2200:1 for each of the five quantitative analyses. As a device for transferring fluid allows you to store and move the fluid within the device, the serial dilution can be carried out inside the system described here, to get these five different dilutions and discover all five different set of analytes, as described here above, and the Protocol for conducting quantitative analyses which also can be adjusted, without the need for modification of the device or system.

In laboratory conditions, the traditional use of pipettes usually take large amounts of sample than in the case of the ROS system. For example, in the laboratory analyses of blood samples taken from the patient's arm in the order of several milliliters. In the case of ROS device when you want the process was quick, easy and minimally inputnum, use small samples (in the order of several microlitres), usually obtained from the fingertip. Due to the difference in volumes of samples, a known ROS devices may lose flexibility in the quantitative analysis, which are laboratory devices. For example, for many quantitative analyses of one sample may require a specified minimum amount for each of the quantitative analysis, in order to perform accurate detection of the analyte, which imposes some restrictions on the ROS device.

In another example described herein, the system and/or device for transferring fluid provide more flexibility. For example, a device for transferring fluid can be automated to move the unit of analysis, tip for quantitative analysis or an empty pipette from one unit into a separate unit, and these units do not have fluid communication drugs other. In some cases, this avoids cross-contamination of blocks of the device. In other cases, it provides the flexibility to move the various fluids within the device that are in contact with each other in accordance with the Protocol or command. For example, when referring to the cartridge, which contains 8 different reagents in 8 different blocks of the reagents may be introduced into the engagement device for transferring fluid, in any order or combination, in accordance with the Protocol. Thus, many different sequences can be used in any chemical reactions in the device. Without changing the volume of reagents in the cartridge or the type of reagents in the cartridge Protocol quantitative analysis can be changed without having a second cartridge or the second system.

For example, a user orders a cartridge with a specific type of surface capture and specific reagents for quantitative analysis, to detect the analyte (for example, C-reactive protein (CRP) in a sample. User Protocol originally planning to conduct a 2 operations of washing and 3 operations dilution. After receiving the user device and the system, the user decided that the Protocol should provide 5 operations of washing and only 1 Opera is July dilution. The proposed devices and systems have the flexibility to change the Protocol without changing the layout of the device or system. In this example, only a new Protocol or set of commands must be sent in a programmable processing device, system or device for transferring fluid.

In another example, the proposed system is configured to conduct five different quantitative analyses of a given analyte, each quantitative analysis should be incubated at different temperatures. In many previously known DFB devices incubation of many quantitative analyses at different temperatures is a difficult task, as many of the quantitative analyses are not modular and the surface of the grip may not move relative to the heating devices. In the system described herein, in which the individual unit of analysis is configured for carrying out a chemical reaction, the individual unit of analysis can be installed in individual heating unit. In accordance with some variations, the system includes a set of blocks of heat. In some cases, the system contains at least as many units of heat as units of analysis. Consequently, many quantitative analyses can be carried out in the centre of the ve temperatures.

Described here are systems and devices also allow the quality control that is missing in many previously known DFB devices. For example, due to the modularity of the device, the quality control units of analysis and units of reagents may be carried out separately from each other and/or separately from the housing and/or separately from the system or device for transferring fluid. The following describes exemplary methods and systems of quality control used in the described systems and devices.

Described here, the system allows a variety of quantitative analyses, regardless of the analyte found in the sample of bodily fluid. The Protocol, which depends on the nature of the device, may be transferred from an external device, where it is stored in the read node device that allows the readout node to use a specific Protocol in the device. In accordance with some variations, the device has an identifier (ID) that is read by the detector ID. The detector ID can be linked to the communication node using the controller, which transmits the identifier to the external device. If necessary, the external device sends, on the basis of the ID of the Protocol that is stored in the external device, the communication node. Protocol, etc the first system, may contain commands to the system controller on the implementation of the Protocol, including (but without limitation) for the specific quantitative analysis and the method of detection. After conducting a quantitative analysis of the system there is a signal that carries information about the analyte in the sample of bodily fluid, which detects the node detection system. The detected signal then through the communication node may be transmitted to an external device for processing, including (but without limitation) to calculate the concentration of the analyte in the sample.

According to some variants, the ID can be the ID of the barcode with a group of black and white lines, which is read by the detector ID, such as a well-known reader barcode. Can be used and other identifiers that contain alphanumeric sequence, the sequence of colors and bumps, and which can be located in the device and can be read using the detector ID. The detector identifier may also be LED, emitting light that is reflected from the ID and is measured in the detector ID to determine the identity of the device. In accordance with some options, identificato who may have a storage device and may transmit the information on the detector ID. In accordance with some options may be used a combination of these means of identification. In accordance with some variations, the detector is calibrated using a light source such as LED.

In accordance with one example, the sample of bodily fluid can be introduced into the device, and the device can be inserted into the system. In accordance with some variations, the device is partially inserted manually, and then a mechanical switch in the read node automatically sets the device to the specified position within the system. However, there may be used other known means of introducing the device or cartridge in the system. In accordance with some variations, the device is inserted into the system manually.

In accordance with some options, we propose a method to automatically select the Protocol executable on the system, which involves the use of a device that contains the detector ID and the ID; the detection of the identifier; transmitting the specified identifier to the external device; and selecting a Protocol executing in the system, many of the protocols in the specified external device associated with the specified ID.

In accordance with the first aspect of the present invention, it is proposed si is theme for the automatic detection of multiple analytes in a sample of bodily fluid, which contains: fluid device (such as described here above), which contains: block sampling, configured to store samples of bodily fluid; matrix units of analysis, and the individual unit of analysis of this matrix blocks analysis is configured for carrying out a chemical reaction, which gives a signal that carries information about the individual analyte specified multiple analytes to be detected; the matrix blocks, reagents, and individual block reagent specified matrix blocks of the reactants contains a reagent. The system further comprises a device for transferring fluid, which includes many heads, and individual head set head configured to enter into engagement with the individual unit of analysis, with the specified device for transferring fluid includes a programmable processing device, configured for direct transfer of the sample of bodily fluid from a block of sample and reagent from the individual unit of the reagent in the individual unit of analysis. For example, the individual unit of analysis contains a reagent and is configured for carrying out chemical reactions with this reagent.

In some cases, the configuration of the processing device, directing the transfer of fluid, allows you to change the power of the ü dilution of the sample of bodily fluid in the matrix units of analysis, to receive signals carrying information on a lot subject to the detection of an analyte within the detection range, so that the specified set of analytes can be detected using this system. In accordance with one example, the sample of bodily fluid contains at least two analyte present at concentrations that differ by at least 2, 5, 10, 15, 50 or 100 orders of magnitude. In accordance with one example, the sample of bodily fluid is a single drop of blood. In one embodiment, the concentration of the at least two analytes present in the sample will differ by 10 orders of magnitude (for example, the first analyte is present at a concentration of 0.1 PG/ml, and the second analyte is present at a concentration of 500 μg/ml In another example, some protein analytes present at concentrations greater than 100 mg/ml, which allows you to expand interest in the range of approximately 12 orders of magnitude.

The degree of dilution of the sample of bodily fluid allows you to receive signals carrying the information of at least two analytes within the detection range. In many cases, the system further comprises a detection device, such as a photomultiplier tube (PMT). In the case of the photomultiplier, the detection range is, for example, approximately from 1 to 10 million counts per second. Each sample corresponds to a single photon. In some cases, the PMT is not 100% effective and the observed velocity calculation is slightly lower than the actual number of photons arriving at the PMT per unit of time. In some cases, the calculations carried out in 10 intervals of approximately one second and the average results. In accordance with some options, ranges for quantitative analyses correspond 1000-1,000,000 counts per second when the device discovery using the PMT. In some cases, the speed of the count may be low and may be 100 times per second, or may be high and may be 10,000,000 times per second. The linear range of response of the PMT (for example, the range in which the speed of calculation is directly proportional to the number of photons per unit time) can be approximately 1000-3,000,000 counts per second. In accordance with one example, a quantitative analysis has a distinct signal of about 200 to 1,000 samples per second at the lower end (range), and of about 10,000 to 2,000,000 times per second at the upper end (range). In some cases, for protein biomarkers speed count is directly proportional to the alkaline phosphatase associated with the engagement surface, and is directly proportional to the analyte concentration. The other item is iminime device detection can be avalanche photodiodes, avalanche photodiode array, CCD and supercooled CCD. Many other detection devices have a digital output signal, which is typically proportional to the number of photons impinging on the detecting device. Detection range sample detecting devices are selected in accordance with the detected analyte.

Individual cylinder device for moving fluid may be configured to interlock with the individual unit of analysis. Device for transferring fluid may be a pipette, such as a pipette with the displacement of air. Device for transferring fluid may be automatic. For example, a device for transferring fluid may further comprise a motor connected to the programmable processing device, and the specified engine can move a lot of heads on the basis of the record from the programmable processor. As described above, the individual unit of analysis may be the tip of a pipette, for example the tip of the pipette with a gripping surface or place of reaction.

Very often, in the DFB device, such as described herein, the system and devices should be considered and it is reasonable to set the dilution factor. For example, in environments in which system employs unskilled users, necessary in order to enforce the dilution of the sample.

As has been described here above, the device for transferring fluid allows you to change the degree of dilution of the sample, in order to give accurate results of the quantitative analysis. For example, a programmable device to transfer fluid can have multiple heads for dilution or serial dilutions of samples and mixing of sample and diluent. Device for transferring fluid also provides the movement of the fluid in the DFB devices.

As has been described here above, the described systems and devices have many of the characteristics of flexibility of laboratory devices in GROWING conditions. For example, samples may be selected and processed automatically in a desktop device or system or to have a smaller size. A common characteristic of ROS devices is to achieve different ranges of dilution at carrying out a variety of quantitative analyses, and quantitative analyses can have very different sensitivity or specificity. For example, in the sample can be two of the analyte, but one analyte has a high concentration in the sample, and the other analyte has a very low concentration. Provided that these systems and devices make it possible to dilute the sample to very different levels, in order to detect both the analysis is as. For example, if the analyte has a high concentration of the sample can be diluted to an appropriate range detection and deposited on the surface of the capture detection. In the same system or in the same device, the sample with the analyte at low concentrations should not be diluted. Thus, widens the range of the quantitative analysis presented here GREW devices and systems compared to many previously known ROS devices.

Device for transferring fluid may be part of a system that represents a table (poster) meter. Device for transferring fluid can have multiple heads. Any number of heads, which is necessary for the detection of multiple analytes in a sample, may be provided in the device to transfer fluid in accordance with the present invention. In accordance with one example, a device for transferring fluid has 8 heads, installed in line with an interval from each other. In one embodiment, the ends are tapered nozzle that using a press fit can be put into engagement with the various tips such as described here, the analysis block or blocks of samples. The tips can be removed automatically by the meter and after use can be stored in the housing of the device is STV. In one embodiment, the tips for the quantitative analysis are transparent and similar ditch, within which perform quantitative analysis, a signal which can be detected using a photodetector such as a photomultiplier.

In accordance with one example, a programmable processor, the system can send commands to the device to transfer fluid for transferring liquid samples, due to the suction or ejection of fluid when the respective piston is moved in a closed air space. As the quantity of air and the movement speed is precisely controlled, for example, using a programmable processor.

Mixing of samples or reagents) with diluents (or other agents) may be carried out by suction mix components in a common tube and then through multiple suction significant fraction of the combined volume of the liquid up and down in the tip. The dissolution of dry reagents in the tube can be carried out similarly. Incubation of liquid samples and reagents with the engagement surface on which is linked to the capture reagent (e.g. antibody), can be achieved through the corresponding suction of liquid into the tip and hold it for a specified time. Destruction of samples and reagents described in the equip can be carried out due to the buoyancy of the liquid in the tank or in the absorbent pad. Then another reagent may be introduced into the handpiece in accordance with the command or Protocol from the programmable controller.

In accordance with one example shown in Fig.11, the liquid 1111, which was previously located at the tip 1101, when it is popping may leave a thin film 1113 inside tip 1101. Therefore, the system uses the first (for example, the upper portions of the following liquid 1112 for washing off the previous fluid 1111 from a tip 1101. Portion 1113 fluid contaminated by the previous liquid can be placed in the upper part of the tip 1101, where it does not interact with the surface 1102 capture. Surface 1102 of the grip can be specified region of the tip 1101 selected so that the previous liquid 1111 does not react with the surface 1102 capture; for example, as shown in Fig.11, the surface 1102 capture occupies a defined area of the cylindrical part of the tip 1101, which does not reach the boss of the tip. In many cases, the incubation period is short (for example, 10 minutes), and contaminated liquid area is relatively long (>1 mm), so that the diffusion of active components contaminated portion 1113 of the liquid does not occur quickly enough to react with the surface 1102 capture during incubation. If m is ogic having high sensitivity quantitative analyses are required removal of one reagent or wash the surface of the capture (for example, in the case of antibody detection (detector), which is observed by the signal generator quantitative analysis). In accordance with one example described herein, the device for transferring fluid system can provide flushing due to the introduction of additional cycles of removal and suction during transfer fluid, for example, using a wash reagent, In accordance with one example, 4 flushing is sufficient for the concentration of unbound antibody detection in contact with the engagement surface was decreased more than 10⁶time. Any antibody that nonspecific associated with the engagement surface (which is highly undesirable), may also be removed during this washing process.

Expansion of the range of quantitative analysis can be achieved by dilution. In DFB analysis systems that use containing diluent disposable cartridges, often there is a practical limit to increasing dilution. For example, if the tip of the finger received a small blood sample (for example, about 20 µl), which must be diluted, and the maximum amount of diluent that can be introduced into the tube, 250 μl, the practical limit dilution of all samples is approximately 10-fold. In accordance with one example, the system can provide is to suck and a smaller volume of sample (e.g., about 2 µl), which allows to obtain approximately 100 times the maximum dilution ratio. For many quantitative analyses of these dilution factors are acceptable, however, in the case of such a quantitative analysis, as CRP (described here in one of the examples), there is a need for further dilution. Based on the division of quantitative ELISA assays are characterized by the restriction of the ability of the surface of the grip to bind the analyte (for example, approximately several hundred ng/ml in the case of a typical protein analyte). Some analytes present in blood at concentrations that comprise hundreds of µg/ml Even at 100-fold dilution of the concentration of the analyte may lie outside the range of calibration. In an exemplary embodiment of the claimed system, the claimed device and the device for transferring fluid may be performed multiple dilutions due to many breaks diluent in the individual unit of analysis, or block sampling. For example, if the concentration of the analyte in the sample is very high, the sample may be diluted several times until the concentration of the analyte will not be within the acceptable range of detection. Described here are systems and methods allow for accurate measurement or estimation of dilutions to RA is to consider the initial concentration of the analyte.

In one embodiment, the proposed system allows you to move the liquid sample and allows you to move the unit of analysis. The system can include a heating unit and a detection unit. To move the liquid sample, the system can perform absorption or action of the action type of syringe or pipette. In an exemplary embodiment, the device for transferring fluid, which moves the liquid sample is a pipette and pipetochnoe head system. The number of devices in the form of a pipette, which is required in a system depends on the type subject to detection of the analyte and the number of conducted quantitative analyses. Action pipettes in the system can be automated or performed manually by the user.

In Fig.5 shows the example described here, the device 520 to transfer fluids and system 500. Device for transferring fluid system can move 8 different or the same amounts of liquid simultaneously, using eight different heads 522. For example, the cartridge (or the described device) 510 contains eight blocks 501 analysis. Individual blocks 501 analysis configured in accordance with the type of quantitative analysis within a block 501. Individual blocks 501 analysis may require a certain volume of the sample. Individual cylinder 522 can be used for the distribution is of the proper amounts of sample in the individual unit 501 analysis. In this example, each cylinder 522 corresponds adisoemarto individual unit 501 of analysis.

The mechanism 520 device to transfer fluid can also be used for distribution of reagents from blocks of reagents. Different types of reagents include the conjugate solution, wash solution and the substrate solution. In the automated system platform 530, shown in Fig.5, on which sits the device 510 may be moved to move the device 510 relative to the location of blocks 501 analysis and heads 522, and in accordance with the operations necessary to perform the quantitative analysis. Alternatively, head 522 and the lugs 501 or device 520 to transfer fluid can be displaced relative to the device 510.

In accordance with some options, reagent receive in dry form and dissolve it in the analysis. Dry forms can be lyophilized material and deposited on the surface of the film.

The system can include a holder for moving blocks of analysis or tips. The holder can be vacuum node or a node that can fit to enter the boss tip for quantitative analysis. For example, the means for moving the lugs can be moved similarly to the head movement device for moving fluid. Device is istwo also can move the platform to the position of the holder.

In one embodiment, the means for moving tips is a similar tool to move the sample volume, such as described here, the device for transferring fluid. For example, the tip of the sampling can fit to enter the cylinder, pipette up to the boss on the tip. Then the tip of the sampling can be used to distribute the fluid across the device and the system. After distribution of the fluid tip sampling can be discarded, and the head of the pipette can be tightly attached to the unit of analysis, to lugs on the unit of analysis. Then the tip for the quantitative analysis may be moved from one block of reagent to another unit of the reagent, and the reagent can be distributed in the unit of analysis due to the action of suction or pipette using a pipette head. The head of the pipette can also produce mixing inside the tip of the sampling, analysis block or block reagent, due to the action of suction or actions of the syringe.

The system can include a heating unit designed to heat the unit of analysis and/or for regulating the temperature of the quantitative analysis. Heat can be used in the operation of the incubation reaction quantitative analysis, in order to accelerate the reaction and to reduce the time required for incubation. The system may have the ü heating unit, configured to receive the unit of analysis in accordance with the present invention. The heating unit may be configured to receive multiple units of analysis device in accordance with the present invention. For example, if 8 of the quantitative analyses, it is desirable to hold the device, the heating unit may be configured to receive 8 units of analysis. In accordance with some options, the units of analysis can be moved (inserted) in thermal contact with the heating block using the moving block analysis. Heating may be effected by means of heat, known by themselves.

Described herein is an exemplary system 600 shown in Fig.6. The system 600 includes a platform 630 translational movement, in which the device 610 (or cartridge in this example) set manually or automatically, or a combination of both. The system 600 also includes block 640 heating, which can be combined with blocks 611 analysis device 610. As shown in Fig.6, the device 610 contains rows of 8 blocks 611 analysis and set of corresponding blocks 612 reagents, and block 640 heating also contains the region 641, in which at least 8 units can be heated simultaneously. In each of the areas 641 heating may be the same or different temperature for each the individual blocks 611 analysis, in accordance with the type conducted a quantitative analysis or a type subject to detection of the analyte. The system 600 also includes a detection device (such as a photomultiplier tube) 650 detection signal from block 611 analysis, which carries information about the analyte in the sample.

In one embodiment, a sensor for localizing unit of analysis relative to the detection device when conducting quantitative analysis.

In one embodiment, the detection device is a card reader, which body is the site of detection for the detection of the signal generated due to at least one of quantitative analysis in the device. Node discovery can be located on the device or may have a different orientation relative to the device, based on, for example, of the type carried out the quantitative analysis and the detection mechanism. Node discovery can be moved to create a connection with the unit of analysis, or the analysis block can be moved to create a connection with the node detection.

In many cases, as the device discovery using the photodetector. As non-restrictive examples of photodetectors include photodiode, photomultiplier tube (PMT) detector photon avalanche photodiode or a charge coupled device (PZ is). In accordance with some variations, can be used in the pin diode. According to some variants, the pin diode may be combined with the amplifier to create a detection device having a sensitivity that is comparable to the sensitivity of the PMT. Some described here quantitative analyses can excite the luminescence. In accordance with some variations, detect chemiluminescence. According to some variants, the node discovery can have multiple fiber-optic cables, the beam of which is connected to a CCD sensor or to the matrix of the PMT. Harness the optical fibers may consist of individual fibers, or many small fibers can be fused together to form a rope. Such harnesses are available for sale, and they are easily joined with CCD sensors.

The detection device may also have a light source such as an incandescent lamp or light emitting diode (LED). The light source used for illumination (venue) quantitative analysis to discover the results. For example, quantitative analysis can be fluorescent quantitative analysis or quantitative absorption analysis that are commonly used in the quantitative analysis of nucleic acids. The detecting device also includes optics for input radiation from the source is the infrared light in the quantitative analysis, such as a lens or fiber optics.

In accordance with some variations, the detection system may be optical sensors for the detection of specific parameters of the subject. Such sensors may be temperature sensors, conductivity, potentiometric sensors amperometric sensors are used to detect oxidized or recovered chemical compounds, such as₂N₂About₂and I₂or oxidized or recovered organic compounds.

The device and system after fabrication can be shipped to the end user, together or individually. The device or system in accordance with the present invention can be packaged together with instructions for use. In one embodiment, the system in accordance with the present invention is common to all types of quantitative analyses carried out in different devices. Because the components of the device may be modular, the user needs only one system and various devices or units of analysis or blocks of reagents for carrying out a variety of quantitative analyses in the place of the care. In this context, the system can be re-used with many devices, however, may require the sensors in the device, and in which the system, to detect, for example, changes in pressure or temperature during shipment. At the time of shipment, changes in pressure or temperature can affect the characteristics of the different components of this system, however, the sensors installed in the device or system, allow us to link these changes, for example, an external device that could be done adjustment during calibration or during data processing to an external device. For example, if the temperature of the fluid device has changed by a certain amount at the time of shipment, the sensor located in the device can detect this change and send the appropriate information to the system when the user inserts the device into the system. To implement these tasks in the system may provide additional detection device (sensor), or the device may be integrated into another component of the system. In accordance with some options, information can be transmitted using radio communication in a system or in an external device such as a personal computer or a display. Similarly installed in the system, the sensor can detect a similar change. In accordance with some options, it is also desirable to have a sensor in the shipping package, instead of the sensor system components or dopolnenie to him. For example, adverse conditions can lead to failure of the cartridge analysis or system, however, this sensor can eliminate the temperature increases above the maximum acceptable level or eliminate the violation of the integrity of the cartridge due to the penetration of moisture.

In one embodiment, the system includes a communication node, which allows you to send and receive information from an external device via wireless communication. In such wireless communication can be used Bluetooth or RTM. Can be used in a variety of ways to communicate, such as a code call using a modem, direct connection, such as T1, ISDN, or cable line. In accordance with some options, your wireless connection is set using such exemplary wireless networks like cellular network, satellite network or subscriber network, GPRS, or a local data transmission system, such as Ethernet or ring network using token-passing access within the local network. In accordance with some options, encode information before its transmission over the wireless network. According to some variants, the communication node may have a wireless infrared communication component for sending and receiving information. The system may have built-in graphics card to facilitate display of information.

Some of the options that, the communication node may be a storage device (memory), for example, localized memory with random access, which can store the collected information. The storage device may be required if the information is not passed in a given time, for example, due to the temporary lack of communication. Information may be combined with the device identifier in the storage device. According to some variants, the communication node may retransmit the stored information after a certain period of time.

In some embodiments, the external device is connected with the communication inside the reader. The external device may be a wireless connection or a physical connection with the system, but may also have a relationship with a third party, including (but without limitation) with the patient, medical staff, clinicians, laboratory personnel, or other individuals involved in the health care field.

An exemplary method and system shown in Fig.7. In accordance with the example shown in Fig.7, the patient enters the blood sample in the described device and then inserts the device into the reader, and the reader may be a desktop system, which allows you to read the analyte in the blood sample. The reader may be in the form of op is pulling systems here. The reader device may be a desktop system, which allows you to read many described here different devices. Reading device or system capable of conducting a chemical reaction and to detect or read the results of chemical reactions. In accordance with the example shown in Fig.7, the reader is automated in accordance with the Protocol, sent from an external device (for example, from the server that contains the user interface). The reader can also send the results of chemical reactions in the server and user interface. In an exemplary system, a user (e.g., a medical worker such as a doctor or researcher) can see the results of the analysis and may accept or to develop a Protocol used to automate the system. The results can also be stored locally (in the reader) or in the server system. The server can also store patient record, register the patient and the patient database.

In Fig.8 shows a block diagram of a system for evaluating a medical condition of the patient. The patient enters personal data and/or measurements from the device described here, the reader device and/or system database, which is described in the server. The system can be scope wirawan for indication of personal data on the display terminal patient. In accordance with some options, the display of the terminal patient is interactive and the patient can change the entered data. The same database or other databases contain data from other patients with a similar medical condition. Data from other patients can be data about the course of the process received from public or private institutions. Data from other patients can be internal data of the clinical examination.

In Fig.8 also illustrates the flow of data from the reader device and entering data into it, which contains patient data passed to the server are connected over a public network. The server can manipulate the data, or simply pass the data to the user terminal. Patient data can also be entered in the server separately, and it can be entered regarding the medical condition that is stored in the database. In Fig.8 also shows the display of the user terminal and the flow of information to medical personnel or user. For example, using the system shown in Fig.8, the patient at home can enter a sample of bodily fluid described herein cartridge in accordance with the present invention and paste it in the system described here or the reader. The patient can see the data obtained is from the system, the display of the terminal patient, and/or may modify the data or enter new data. Then the data from the patient pass through the public network such as the Internet, for example, in coded form, and enter the server that contains the network interface and the processor, and the server is located in the computer center or in the clinical research center. The server can use these medical condition for processing and understanding received from the user data, and can then send the results on the public network to the user terminal. The user terminal may be located in the medical office or in the laboratory, and may have a display to indicate the results of the data analysis and processing of the patient by medical personnel. In this example, the medical staff can get the results and analysis of the sample from the patient obtained through tests that the patient is held in a different location, for example at home. Can be used and other options and examples described herein systems and components of systems.

In accordance with some variations, an external device may be a computer system, server, or other electronic device that can store information or to process information. In accordance with some variations, an external device sod is rgit one or more computer systems, one or more servers or one or more other electronic devices that can store information or to process information. In accordance with some variations, an external device may have a database with information regarding a patient, for example (but without limitation) with medical records or patient history, with records of clinical trials or with records of animal testing. The external device can store protocols running in the system, which may be transmitted in a communication system, when he received identifier that indicates which device has been inserted into the system. According to some variants, the Protocol may depend on the device ID. According to some variants, the external device stores multiple protocols for each device. In accordance with other options, information regarding a patient external device includes multiple protocols. In some cases, the external server stores the mathematical algorithms for processing times of the photons received from the communication node, and uses, in accordance with some options, mathematical algorithms to calculate the analyte concentration in the sample of bodily fluid.

In accordance with some options, external devices which may have one or more of the purchase of servers, as is well known to experts in this field. Each server can provide load balancing, task management and has the capacity of the backup memory, which is used in case of failure of one or more servers or other components of the external device, to increase the availability of the server. The server can also be implemented as a distributed network block storage and processing of information, which in itself is known, and the processing of data in accordance with the present invention produce on workstations such as the computers, which eliminates the need to use separate server.

The server may have a database and can be subjected to the data processing system. The database can reside within a given server or may reside in another server system has access to this server. Since the information in the database may contain sensitive information, may be provided with a security system that does not allow unregistered users to have access to the database.

One of the advantages of some of the characteristics described here is that information can be transferred from the external device back, not only in the host reading device, but in other parts of the system or in others the many external devices, including, for example (but without limitation) in the "pocket computer (PDA) or cell phone. Some types of communication can be implemented using a network with a wireless connection, as mentioned here above. According to some variants, the calculated concentration of the analyte or other related patient information can be sent, for example (but without limitation) to medical personnel or the patient.

Thus, the data obtained by the claimed devices and systems may be used to perform a trend analysis of the concentration of analyte in a patient.

Another advantage of the present invention is that the results of the quantitative analysis can be immediately transferred to any third party who may benefit from the results. For example, after determining the analyte concentration in the external device, it can be transferred to the patient or to medical personnel, which may take additional action. The operation of transmitting information to a third party may be implemented using wireless communication, as mentioned here above, for example, by transferring data to a mobile device of a third party so the third party may receive the results of the quantitative analysis is actually at any time is anywhere. Thus, in scenarios with lack of time patient contact may be carried out immediately, if you need immediate medical action.

Due to the detection device based on the identifier associated with the fluid device after its introduction into the system, the system allows you to download specific fluidic device protocols from an external device and to fulfill them. According to some variants, the external device can store multiple protocols, system-related or related to a specific patient or group of patients. For example, when the ID is passed for connection to an external device, the program in the external device can obtain the ID. After receiving the identifier, the software in the external device, such as a database, can use the ID to identify protocols that are stored in the database associated with the ID. If ID is associated with only one Protocol, then, for example, the database can choose the Protocol and software in the external device and then can transmit the Protocol in a communication system. The use of protocols, a specific image associated with the device, allows you to use any component of the device in accordance with the present the invention in a single system, so in fact, any interest of the analyte can be detected in a single system.

In accordance with some variations, multiple protocols can be associated with a single identifier. For example, if is the preferred detection of a single analyte of the patient once a week, and the other of the analyte from the same patient twice a week, the protocols in the external device associated with the identifier can also be associated with different days of the week, so when the identifier is detected, the program in the external device can select an appropriate Protocol that is associated with a specific day of the week.

In accordance with some options, the patient can have many of the claimed devices used to detect different analytes. The patient, for example, may use different devices in different days of the week. In accordance with some options, the program in the external device, linking the identifier to the Protocol may include the process of comparing the current day to day use of the device, for example, on the basis of clinical examination. If, for example, two days of the week are not the same, the external device can use wireless communication to notify the patient, using any of the described method is in or other known methods, that wrong device inserted in the system, but also about what the correct device you want to use that day. This example is only explanatory and can easily be extended, for example, when notify the patient that the device was not used at the right time of day.

The system can also be used the method of connection to the network for evaluating a medical condition of the patient. The communication system may or may not have a reader for reading data of the patient. For example, if the biomarker data obtained using microfluidic GREW device, then the indicators associated with various individual biomarkers that can be read by means of this device or another separate device. Another example of a reader is a system with a bar code that allows you to scan patient data that were entered in the electronic medical record or map of therapist. Another example of a reader database is the electronic patient record, from which patient data can be directly obtained by using the communication network. Currently, the effectiveness of specific drugs can be evaluated in real time, allowing you to justify the expense of those who Apia.

Failure to comply with treatment regimens, including defined through clinical examination, can seriously reduce the effectiveness of treatment. Therefore, in accordance with some options, the claimed system may be used to monitor adherence to the treatment by the patient and notify the patient or medical personnel about the failure modes. For example, if a patient who takes the drug that is part of the plan of medical treatment, take samples of bodily fluids to conduct described herein quantitative analysis, and analysis system detects, for example, elevated concentrations of the metabolite compared to the known profile, this may indicate an (unauthorized) taking multiple doses of the drug. The patient or medical personnel can be notified of such failure to comply with treatment regimens by means of wireless communication, for example, including (but without limitation) using mobile devices such as pocket PC or mobile phone. Specified a known profile may be stored in this external storage device.

In one embodiment, the system can be used to identify subpopulations of patients for whom therapy is favorable or harmful. For accounts is this medication with varying toxicity, commercially available, will be assigned only to those patients for whom they are not harmful.

Ways

The devices and methods in accordance with the present invention provide an effective means of detection in real time of analytes present in a bodily fluid of the patient. The claimed detection methods can be used in a variety of circumstances, including identification and quantification (quantitative determination) of analytes that are associated with specific biological processes, physiological conditions, disorders (disorders) disorders or stages, or phases of therapy. As such, these devices and methods have a wide range of applications, for example, for drug screening, diagnosis, phylogenetic classification, paternity and forensic identification, the beginning and repeat diseases, individual response to the treatment of different population groups and for monitoring therapy. Offered here is a device and system is also particularly useful for the assessment of preclinical and clinical stages of therapy, to improve adherence to medical treatment of a patient, monitoring of erythrocytes associated with the designated drug for the development of individualized medicine that provides the translated review of blood from the Central laboratory to the test at home of the patient. The device can also be used on the basis of a prescription (prescription basis) used by pharmaceutical companies to monitor therapeutic agents after receiving permission for their use.Thus, in accordance with one variant of the present invention, it is proposed a method for detecting analyte in a sample of bodily fluid, which provides for the introduction of (flow) of the blood sample into the device or system in accordance with the present invention, creation of conditions for response specified sample within at least one unit of analysis, and detection of a specified audible signal generated by the specified analyte in the blood sample.

In Fig.1 shows an exemplary variant of the device in accordance with the present invention, which contains at least one analysis block and at least one unit of the reagent. The units of analysis (e.g., as the tips of sampling and tips for the calibration of Fig.1) may have a gripping surface, and blocks reagents may contain tools such as conjugates, wash solutions and substrates. The exemplary device shown in Fig.1, also includes a tip for collecting a sample of whole blood, the tip for the selection of blood plasma, the hole for receiving the blood, the hole for the separation of blood plasma, the tip to drain or vetiva the General (hygroscopic) gasket, well for dilution, reagent diluted blood plasma or the reagent diluent blood plasma, as well as the storage area of the exhaust tips.

In one embodiment, the method involves the implementation of quantitative analysis with an associated enzyme immunosorbent assay (ELISA). In accordance with one example described in this paragraph, the sample is fed into the block sampling device described here. The device is then inserted into the system, and the system detects the type of the entered device or cartridge. The system can then communicate with an external device for receiving command set or Protocol that allows the system to carry out the desired quantitative analysis or quantitative analyses of the cartridge (the contents of the cartridge). The Protocol can be sent to a programmable processor device for transferring fluid system. In accordance with one example, a device for transferring fluid engages with the tip of the sample cartridge, selects a certain volume of sample of the block sampling and moves it in the preprocessing block, which removed the red blood cells. Can then be made sucking the sample of blood plasma in the tip for blood plasma or in another tip, for quantitative analysis, using the device for transferring fluid, in accordance with the Protocol. N is konecnik, containing the blood plasma, then there may be a diluent to dilute the sample in accordance with the conducted quantitative analyses. Many different dilutions can be performed through the use of serial dilutions of the sample. For example, each tip for quantitative analysis or analysis block may contain samples with different dilutions. After absorption of the sample in the unit of analysis, using the device for transferring fluid, the unit of analysis can then be incubated breakdown to create conditions for the attachment of any present given analyte to the surface of the grip. Described in this example, incubation can be carried out at room temperature for a predetermined period of time, for example, within 10 minutes, or can be conducted by heating described here, the devices of the system. The unit of analysis can be put into engagement with the block reagent containing the reagent corresponding to the conducted quantitative analysis of each individual unit of analysis, which has a gripping surface for such a quantitative analysis. In this example, the first reagent is a reagent for detection in the analysis, ELISA, for example, which contains the antibody, such as a labeled anti-protein antibody that differs from the surface of the grip. Rast is the PR for detection then removed from the block analysis and suck the wash solution in the unit of analysis to remove any excess detection solution. Can be used for many operations rinsing. The final added reagent is a substrate of the enzyme, which produces a binding solution for detection by chemiluminescence. Then, the substrate of the enzyme is removed from the unit of analysis and the results of the analysis are read using device detection system. In each of the operations described here can be done is described incubation. In this example, the whole process after you install the cartridge into the system is automatic and takes place in accordance with the Protocol or set of commands entered into the programmable system.

One exemplary method begins by entering the blood samples in the hole for blood. The sample can then be selected tip for the selection and put in the hole for separating the blood plasma. Alternatively, blood can be directly introduced into the hole, which contains the blood separator. For example, the separation of blood plasma can be carried out using various methods described here. In this example, the separation of blood plasma can be performed using magnetic beads and antibodies to remove blood components that are not blood plasma. The plasma can then be transferred using a handpiece selection of blood plasma, so as not to contaminate a sample of whole blood from the tip of the selection of CROs is I. In this example, the tip of the sampling of blood plasma can select a specified number of diluent to the dilution of blood plasma. Diluted sample of blood plasma distributed in blocks (tips) analysis to associate with the engagement surface. The units of analysis can be incubated for creating conditions for reaction on the surface of the grip. The unit of analysis can then be used to collect the conjugate to bind to the reaction unit of analysis. The conjugate can have a part that allows you to discover the interest of the analyte using a detection device, such as a photodetector. After adding a conjugate to the unit of analysis, the reaction can be incubated. In an exemplary method of using an exemplary device shown in Fig.1, a block reagent containing wash solution to conjugate, then enter into contact with the unit of analysis to remove any excess conjugate, which can interfere with the detection of the analyte. After washing the excess conjugate, the unit of analysis can be added to the substrate for detection. In addition, in accordance with one example shown in Fig.1, this method can be used block (NIB) calibration, which is used in all described in this paragraph operations, except for the selection and allocation of samples is. Detection and measurement using block calibration can be used to calibrate the detection and measurement of the analyte in the sample. The following describes other processes and methods similar to those described in this example.

Any bodily fluid that may contain the interest of the analyte, can be used to analyze a system or device in accordance with the present invention. For example, in the input hole or block sampling in accordance with one example shown in Fig.1, it is possible to collect and store any type of commonly used bodily fluids, including (but not limited to, blood, serum, saliva, urine, gastric and digestion fluids, tears, feces, semen, vaginal fluid, interstitial fluid, obtained from tumor tissue, and cerebrospinal fluid. In one embodiment, the bodily fluid is blood, which can be selected from the fingertip. In one embodiment, the sample of bodily fluid is a sample of blood plasma.

The bodily fluid may be withdrawn from the patient and dispensed into the device through a variety of means, including (but without limitation) using tubes, injections or pipette. In one embodiment, the Lancet pierces the skin and takes samples using, for example, gravity, capillary the th action suction or vacuum. The Lancet may be part of a device or part of a system, or a separate component. If necessary, the Lancet can be powered using a variety of mechanical, electrical, Electromechanical or any other known mechanism actuation, or any combination thereof. In another embodiment, there is no active mechanism is not required, as the patient can enter bodily fluid in the device, for example, when introducing a sample of saliva. Collected fluid may be placed into the hole or block and collect the sample device. In accordance with some options, the user actuates the Lancet and capillary tube to collect the sample within the device.

The volume of bodily fluid used in the here described method or device, usually is approximately less than 500 μl, and may be approximately from 1 to 100 μl. If necessary, the sample is from 1 to 50 μl, 1 to 40 μl, 1 to 30 μl, 1 to 10 μl, or even from 1 to 3 ál can be used for detection of the analyte using the claimed fluid device. In one embodiment, the volume of the sample is 20 ál.

In one embodiment, the volume of bodily fluid, used for detection of the analyte using the claimed devices, systems or methods, is equal to the volume of a single drop of liquid. the example one drop of blood from a pricked finger may form a sample of bodily fluid analysis device, system or method in accordance with the present invention.

In accordance with some variations, the body fluids are used directly for the detection of analytes present in a bodily fluid, without additional processing. However, if necessary, can be carried out pre-processing of bodily fluids to the analysis device. The choice of the form of pre-treatment depends on the type used bodily fluids and/or investigational nature of the analyte. For example, when the analyte is present at low levels in a sample of bodily fluid, the sample may be concentrated by any conventional means to increase the concentration of the analyte. Ways of increasing the concentration of the analyte include, but are not limited to, drying, evaporation, centrifugation, sedimentation, sedimentation and amplification. When the analyte is nucleic acid, it can be extracted using various lytic enzymes or chemical solutions, or using polymers for binding nucleic acid, in accordance with the instructions of the manufacturers. When the analyte is a molecule on the surface or inside the cell,the extraction can be carried out using various lytic enzymes, including (but without limitation) anticoagulants, such as ethylenediaminetetraacetate or heparin, denatured detergent, such as SDS, or not denatured detergent, such as Thesit, deoxit sodium, Triton X-100 and tween-20.

In one embodiment, the patient selects a sample of bodily fluid using a syringe. The sample can be introduced into the syringe through the capillary (capillary) tube. In the embodiment, measurement of the analyte in the blood sample, the patient pierces the tip of your finger and touches the outer edge of the glass capillary tube so as to produce a suction of blood through capillary action and enter the required volume of blood in the capillary tube. In some cases, the volume of the sample is known. In accordance with some variations, the volume of the sample lies in the range of approximately 5-20 µl, or it may be another suitable range.

In another embodiment, the method and system is used to obtain samples of blood plasma mainly containing erythrocytes. When carrying out the quantitative analysis, the analytes are often contained in the blood plasma, and red blood cells may interfere with the reaction.

When measuring blood samples of interest, the analytes are often found in the serum or in plasma. For clinical purposes, it is often necessary to link the final concentration of the many trials of blood concentration saw rocky blood or blood plasma in the diluted sample. In many cases, serum or plasma are the preferred control environment in the laboratory. Two operations may be required earlier quantitative analysis, namely, the dilution of the sample and destruction of erythrocytes. Blood samples may be significantly different volume occupied by erythrocytes (changing hematocrit approximately 20-60%). Moreover, in the device at the site of patient care, when the analysis system uses non-qualified personnel, the resulting sample may not correspond to the required volume. If you do not change this amount, it may cause error in the measured concentrations of analytes.

In accordance with a related but separate embodiment, in accordance with the present invention it is proposed a method of sampling blood plasma from blood samples, which provides for mixing of blood samples in the presence of magnetized particles in the unit of sampling, and the magnetized particles contain surface of the capture antibody to bind not plasma portions of the blood sample, and the imposition of a magnetic field on top of the collection zone of the plasma, for mixing blood samples, to obtain a suspension plasma portions of the blood sample on top of the collection zone of the plasma, thereby deprived of the plasma from the blood samples.

When the blood is mixed with a magnetic reagent, can occur together agglutination, in which many, if not all, of the red blood cells form a mixed agglutinate with the magnetized particles. The process of dissolution and mixing of the reagent is carried out through multiple suction using a handpiece in accordance with the present invention or tip in the form of a pipette. After the formation of the magnetized mass, this mass can be separated from the blood plasma by use of a magnet, which holds the weight in place at the time of blood plasma from the tip. In one embodiment, the blood plasma out of the tip by gravity in the vertical direction, while the magnet holds the weight in place. In another embodiment, the blood plasma out of the tip by application of vacuum or pressure, until the mixture is kept inside the tip. The blood plasma may be introduced into the hole in the other of the tip or in the unit of analysis in accordance with the present invention.

An example of a method of separating blood plasma in accordance with the present invention shown in Fig.9A-9F. In Fig.9A shows that the sample 901 whole blood suck in this tip 910 sampling, for example, of about 20 ml. Then, the sample 901 whole blood is introduced into the separating hole 920 (e.g., in the hole, which is contains magnetic beads or particles) approximate unit. In Fig.9B shows how the suspension and mixing of magnetic reagent in the sample 902 whole blood into the separation hole (for example, a reagent containing the circular magnetic particles and free of binding molecules). In Fig.9C shows an air tube 930 volume of 10 μl, which can be used to prevent loss of material from the tip 910. The mixture 902 samples of whole blood and the magnetic reagent is incubated for several seconds (for example, from 60 to 180 seconds) to perform the agglutination reaction.

In Fig.9D shows the application of a magnetic field 940 to the mixture 902 samples of whole blood and the magnetic reagent. Magnetic field 940 may be applied by means of a magnetic ring 942 built into the system, or by other known magnetic means. Magnetic field 940 attract any particles that are stuck to the magnetic reagent. Due to this, plasma 903 blood, which does not stick to a magnetic reagent, can be separated from plasma sample portions of whole blood.

In Fig.9F shows the distribution of the sample 903 blood plasma, obtained by the described separation due to a magnetic reagent, in the hole or block 950 described device. Sample 903 plasma can also be entered in the tip or the unit of analysis, as well as in any other part of the device for the quantitative analysis, Thu is known to experts in this field. In Fig.9F shows the magnetic field 940, which moves with the tip 910 in the distribution of the sample 903 blood plasma. In this example, select 5 to 8 µl of blood plasma from a 20 µl sample of whole blood. Using the method of separation of blood plasma in accordance with the present invention, from 1 to 99% of a sample of whole blood to form a blood plasma, which can be separated. In one embodiment, from 25 to 60% of the sample volume of whole blood to form a blood plasma, which can be separated.

To complete the described method can be used in other exemplary operations. To move samples of blood plasma in the other hole or other unit, can be used capillary tip of the collection of blood plasma (which can be controlled using a robotic system or any other system in accordance with the present invention) in order to collect a sample of blood plasma due to capillary action and suction power. Another operation may be dilution of plasma samples blood diluent. A specified volume of diluted sample of blood plasma can then be selected with the appropriate tip. Then, a diluted sample of blood plasma can be mixed and introduced into the hole or block device, for distribution to one or many blocks of the analysis device in accordance with the present invention the Sample may also be distributed in other types of device, such as titrations the microplate, as is well known to experts in this field.

The exemplary method shown in Fig.9A-9F may be used with other devices and systems, which differ from those described here. For example, the tip to transfer fluid may contain agglutinating mass, and plasma may be in titrations the microplate. Other known devices and systems, as is well known to experts in this field can also be used to effect the separation of blood plasma.

The sample of bodily fluid can also be diluted in any other way, for example, using a device for sampling that allows for dilution. The device for sampling can have the camera. In this chamber, between two movable sealing gaskets, may be the volume of diluent. In accordance with the preferred option, the amount of diluent is specified, for example, is approximately from 50 μl to 1 ml, and mostly approximately from 100 μl to 500 μl.

In accordance with another aspect of the present invention, it is proposed a method of automatic detection of multiple analytes in a sample of bodily fluid, which includes the following: introduction of the sample of bodily fluid in fluid the e device moreover, the fluidic device includes: a unit of sampling, configured to store samples of bodily fluid; matrix units of analysis, and the individual unit of analysis of this matrix blocks analysis is configured for carrying out a chemical reaction, which gives a signal that carries information about the individual analyte specified set of detectable analytes; and the matrix blocks, reagents, and individual block reagent specified matrix blocks of the reactants contains a reagent. The method may also involve the introduction engages the individual unit of analysis using a device for transferring fluid. To continue the process, the sample of bodily fluid can be transferred from a block of samples in the individual unit of analysis using the device to transfer fluid and the reagent from the individual unit of the reagent can be transferred to the individual unit of analysis, resulting in reaction of the reagent with a sample of bodily fluid that gives a signal that carries information about the individual analyte many to be the detection of analytes. According to some variations, a device for transferring fluid contains many heads, and individual head set head configured to enter into engagement with the individual unit of analysis;however, the specified device for transferring fluid includes a programmable processing device, configured for direct transfer fluid sample of bodily fluid from a block of sample and reagent from the individual unit of the reagent in the individual unit of analysis.

In some cases, the commands in the programmable processing device specifies, for example, the user, the patient or the manufacturer. Commands can be received from an external device, such as a personal electronic device or server. The team can control the transfer operation of the sample of bodily fluid in the individual unit of analysis. For example, the transfer operation of the sample of bodily fluid can alter the degree of dilution of the sample of bodily fluid in the individual unit of analysis, in order to obtain a signal that carries information about an individual analyte of multiple analytes to be detected within the detection range. In some examples, the degree of dilution of the sample of bodily fluid allows you to receive signals carrying the information of at least two individual analytes within the detection range.

The technique of pattern recognition can be used to determine that the detection of the analyte or multiple analytes using the method described here produced within a specified range or outside it. For example, can be discarded distinguishable signals, if they are ahadada outside the specified range. The specified range can be formed in the calibration fluid device, and blocks of reagents and unit of analysis. For example, the range can be specified at Assembly and setup (just-in-tune fashion).

In some cases, if visible signal of the analyte detected at a lower dilution ratio or dilution exceeds the signal at higher dilution factor, then the result obtained at a lower dilution ratio should be discarded as invalid. In most cases, the concentration of the analyte in the samples with different dilution factors give signals, which decrease with increasing dilution. If not, the results of the quantitative analysis should be checked. Described here are systems, devices and methods provide the flexibility to conduct quality control which is lacking in many previously known ROS devices. Described here are systems, devices and methods have many of the characteristics that are required in laboratory devices.

In one embodiment, the sample is diluted in such a ratio that is suitable for quantitative analyses of both high and low sensitivity. For example, the dilution ratio of the sample diluent may lie in the approximate range of 1:10,000-1:1 Device allows for dilution of the sample in different places or in different ways. The device also allows a serial dilution of the sample. In accordance with other options, a serial dilution within the device or system allows to dilute the sample to 10,000,000,000:1.

In one embodiment, contains detectable analyte sample can be moved from the first location to the second location due to the absorption or action of the syringe or pipette. The sample may be drawn into the reaction tip through capillary action or vacuum to atmospheric pressure. According to some variants, the sample is moved to different locations, including in the matrix blocks of the analysis device in accordance with the present invention and various holes in the device in accordance with the present invention. The process of moving samples can be automated using the system described here in accordance with the present invention.

The units of analysis and/or containing a sample of the tips can also be moved from the first location to the second location. The process of moving block analysis or the tip of the sampling can be automated and can be carried out using a user-specified Protocol.

In one embodiment, the blocks move analysis for the selection of the reagent from a block of real the NTA in accordance with the present invention. In many cases, the moving block analysis can be automated. The selection of the reagent from a block of reagent in the unit of analysis can be carried out by absorption or action of the syringe or pipette.

After entering the sample in the analysis block, which contains the engagement surface, the entire unit may be incubated for a specified period of time, to create the conditions for the reaction between the sample and the surface of the capture unit of analysis. The period of time required for incubation of the reaction often depends on the type conducted quantitative analysis. The process can be automated using a system in accordance with the present invention. In one embodiment, the incubation time is from 30 seconds to 60 minutes. In another embodiment, the incubation time is 10 minutes.

The unit of analysis can also be incubated at an elevated temperature. In one embodiment, the unit of analysis is incubated at a temperature in the range of approximately 20 to 70 degrees Celsius. The unit of analysis may be inserted into the heating unit to raise the temperature of unit of analysis and/or the temperature of the contents of the unit of analysis.

In a variant of the method in accordance with the present invention, the conjugate is added to the unit of analysis after the introduction of the sample into the unit of analysis. The conjugate may be of the molecule to be used for labeling the analyte captured by the surface to which the unit of analysis. Here are examples of conjugates and surface grip. The conjugate may be a reagent, which is located within a block of the reagent. The conjugate can be distributed in the unit of analysis due to the absorption or action of the syringe or pipette. After entering the conjugate in the unit of analysis the unit of analysis can be incubated in order to create conditions for the reaction of the conjugate with the analyte inside the unit of analysis. The incubation period depends on the type conducted quantitative analysis or the type should be detected analyte. The incubation temperature may be any temperature necessary for reaction.

In accordance with another aspect of the present invention, it is proposed a method of calibrating the device for automatic detection of analyte in a sample of bodily fluid. The device may have a matrix of addressable units of analysis that are configured for carrying out a chemical reaction, which gives a distinct signal that carries information about the presence or absence of the analyte and the matrix addressable blocks of reagents, which apply to each of the blocks of reagents corresponding to one or more addressable units of analysis in the specified device, so that the individual blocks of the reagents are calibrated by a reference signal of the corresponding block (block) analysis, previously the Assembly of the matrices in the device. Device Caleb the coziness by means of calibration units of analysis and units of reagents previously installed in the device. The device can then be collected using a calibrated components, making the device, and method and system using the device of modular components.

The calibration can be performed using preliminary measurements of the characteristics of the reagents for the quantitative analysis, such as conjugates, previously Assembly units of analysis and the unit of the reagent in the device in accordance with the present invention. Information and calibration algorithms can be stored in a server connected via wireless to the system for the quantitative analysis. The calibration can be made in advance or retrospectively, at the expense of the quantitative analyses carried out in the copies of the system in a separate location, or through the use of information obtained during use of the system for the quantitative analysis.

In one aspect, the control material can be used in the device or system for measuring or checking the degree of dilution of the sample of bodily fluid. For example, quantitative analysis based on a solid phase, such as ELISA, are a quantitative analysis, which uses the reagent in the solid phase, quality control, which is difficult without destroying its function. Described here are systems and methods allow us to determine rubbable is s, received GREW up in the system, with the use of disposable devices with automated mixing and/or dilution.

In one embodiment, the method involves retrospective analysis, for example, through the use of the server in real time for analysis of data previously obtained results. For example, quantitative analysis and control quantitative analysis can be carried out simultaneously. Control quantitative analysis allows measurement of the expected dilution. In some examples, control, quantitative analysis allows you to check the dilution of the sample and, thus, dilution of the sample for the quantitative analysis or multiple quantitative analyses carried out within the system, which can be regarded as precise.

A method of measuring the volume of the liquid sample may include: dealing with a known quantity of a control analyte in the liquid sample with a reagent to obtain a visible signal that carries information about the control analyte; comparing the intensity of the specified visible signal with the expected intensity of a specified audible signal, and the expected intensity of the specified signal carries information about the expected volume of the liquid sample, thus this comparison makes it possible to measure the indicated amount is as specified liquid samples. In many cases, the control analyte is not present in the specified liquid sample in a detectable quantity.

In one embodiment, the method may further include checking the specified volume of the liquid sample when the measurement results of the sample volume are approximately less than 50% of the expected volume of the liquid sample.

For example, a method in which use is described here, the device or system may further include: response of the sample of bodily fluid containing the specified analyte, reagent, to obtain a visible signal that carries information about a given analyte; and measuring the amount of a given analyte in a sample of bodily fluid using the specified intensity audible signal carrying information about a given analyte, and measuring the specified amount of the specified liquid samples. Liquid sample and the breakdown of bodily fluids can be the same sample. In accordance with some variations, the control analyte does not react with a given analyte in a sample of bodily fluid, so that there is no interaction in the detection of a given analyte.

In some cases, the liquid sample and the sample of bodily fluid represent different samples. For example, the control fluid is water, and a breakdown of bodily fluid is a blood sample or, in another example, trolley fluid is a sample of saliva, and a breakdown of bodily fluid is a blood sample.

The control analyte may be (but not limited to, fluorescein labeled albumin labeled with fluorescein IgG, anti-fluorescein, anti-digoxigenin labeled digoxigenin albumin, labeled digoxigenin IgG, biotinylated proteins and IgG is not human. Specialists in this area and other known approximate control analytes. In one embodiment, the control analyte contained in a sample of bodily fluid of a person.

Described here GREW up system, configured to detect multiple analytes within a sample, you may have the opportunity for dilution and mixing of liquids. In many cases, the automated system allows you to use or the user can use the control quantitative analysis to measure really made of dilution and the dilution factor in the calibration system. For example, the control analyte may never be detected in the interest of the sample and dried in block reagent. A known amount of dried control analyte may be mixed with the sample in the block reagent. The concentration of the analyte can be measured to determine the volume of the sample and conducted any dilution of the sample.

As examples of control analytes for immunological the analysis can lead to (but without limitation): fluorescein labeled protein, biotinylation protein labeled with fluorescein-labeled Axlexa™, labeled with rhodamine-labeled red pigment Texas Red, immunoglobulin. For example, the marking can be achieved through at least two haptens that are associated with each molecule of protein. In accordance with some options, 1-20 haptens bound to each protein molecule. In accordance with another option, 4-10 haptens bound to each protein molecule. Many proteins have a large number of free amino groups which can be bound haptens. In many cases, modified by haptens proteins are stable and soluble. In addition, such haptens as fluorescein and red pigment Texas Red are sufficiently large and rigid and allow you to create antibodies with high affinity (for example, the hapten is large enough to fill the binding site of antibodies). In accordance with some options, haptens can be attached to proteins using reagents such as isothiocyanate fluorescein and ether NHS fluorescein carboxylic acid to generate the control analytes, which are identifiable by means of the quantitative analysis part is a hapten.

In accordance with some options in the way they use dried the control analyte. In some examples, izuchenii control analyte avoids dilution of the sample, what makes the control analyte is more stable. The dried composition of the control analyte may be selected so that it is rapidly and/or completely dissolves when exposed to the liquid sample. In accordance with some variations, the control analyte may be an analyte with high affinity to antibodies. In some cases, the control analyte may be the analyte, which has no adverse reactions with any of the endogenous component of the sample. In addition, the analyte is cheap and/or easy to manufacture. In accordance with some variations, the control analyte is stable for the life described here, the device or system. As an example of media used to create analytes with covalently linked heptane, can result in proteins, such as (but not limited to, albumin, IgG and casein. As exemplary polymeric carriers used to create new analytes with covalently linked heptane, can result in (but not limited to, dextran and polyvinylpyrrolidone. As an example of the fillers used for the formation and stabilization control of an analyte that can result in (but not limited to, sucrose, salt and buffer solutions (such as sodium phosphate and Tris chloride).

Described here control analyte and method of calibration can be is used in different ways, including in accordance with the examples described here. For example, the method allows to measure the volume of the sample. In accordance with some variations, the method allows to measure the dilution or dilution ratio or the degree of dilution. In some cases, the method allows to measure the concentration of control of the analyte in the sample. In the system described herein or device for detecting a variety of analytes, measurement in this way using control analyte can be used to validate measurements of specified analytes. For example, a device for transferring fluid with many heads can be used to distribute the fluid in multiple units of analysis, including in the control unit. In some instances, it is believed that, when the distribution of fluid in many units, individual units receive the same amount of liquid. According to some variants, the described method validation with the control analyte may be used to check what was selected or used the correct volume of sample within the device or system. In another embodiment, the method may be used to check the amount of diluent introduced into the sample. In addition, can be checked the dilution ratio or the degree of dilution. In accordance with the one school option method validation with the control analyte may be used to verify that the correct volume of sample was divided into many blocks.

In Fig.10 shows an exemplary method is described here control the quantitative analysis, which use known number of control analyte. Block 1010 earlier introduction (Assembly) in the cartridge can be filled with a solution 1001, which contains a known weight of the control analyte 1002. The liquid solution may be removed and the block 1010 dried to leave the control analyte 1002 in block 1010. Block 1010 may then be inserted into the device and intended for use. When a block 1010 is used and injected in the sample or diluent 1003 in the expected number, the sample 1003 mixed with dried control analyte 1002 within a block 1010 to generate a control solution 1004 with the expected concentration. The control solution 1004 may optionally be diluted. In one embodiment, the control analyte 1002 can be detected similarly to the detection of a given analyte in the device. Measure the concentration of control of the analyte in the test solution 1004. The concentration measurement can be used to calculate the sample volume 1003, added to create a control solution 1004. Thus, the user can compare the measured volume of sample is expected sample volume 1003.

In accordance with one example, the erythrocytes can be removed from blood samples. However, if some portion of the erythrocytes remained or if the red blood cells are not removed from the blood samples, the method with the control analyte may be used to adjust the influence of erythrocytes in the blood sample. As the hematocrit can vary greatly (for example, from 20-60% of the total sample volume), the amount of analyte in a fixed or expected volume (v) of blood may be a function of hematocrit (H is expressed here as a decimal fraction). For example, the number of analyte concentration in blood plasma is C*v*(l-H). Thus, the number of samples with hematocrit 0.3 1.4 times greater than the number of samples with hematocrit 0.5. In an exemplary embodiment, the undiluted blood can be distributed in the device described here and erythrocytes can be removed. Then can be measured the concentration of control of the analyte in plasma fractions to estimate the amount of samples of blood plasma and to determine the hematocrit.

In accordance with some options, you may want to wash away the unbound conjugate from the reaction, to prevent the wrong detection of the unbound conjugates. Limiting operation of many immunological assays is the washing operation. A little compromise on migration and high chustvitelnos and depends on the removal by washing the unbound conjugate. The washing operation may be severely limited in the format tiralongo microplate because of the difficulty of removing washing liquid from the wells (e.g., using automated means). The unit of analysis devices and systems in accordance with the present invention has several advantages in the treatment of liquids. One of the benefits is to increase the signal-to - noise quantitative analysis.

Removal of the conjugate can be difficult if the conjugates stick to the edges of the blocks of the analysis device and when, for example, there is no excess wash solution.

Flushing conjugate can be made by the discharge of the washing solution from the top or suction of the washing solution up and removal of fluid similar to loading samples. Rinse if necessary, can be repeated several times.

When in quantitative analysis using wash buffer solution, the device can store wash buffer solution in blocks of reagents, and the unit of analysis can be put in fluid communication with the flushing. In one embodiment, the wash reagent allows you to remove by washing approximately up to 99.99.9 or 99.999% of the unbound reagent of the units of analysis. As a rule, preferred is a high wash ability, allowing to obtain a high degree of reduction of the possible unwanted background signals. Washing efficiency, which is typically defined as the ratio of the signal from the quantitative analysis to the full value of the signal obtained by the quantitative analysis without flushing, can be easily determined using standard experiments. Usually it is preferred to increase the volume of wash solution and the incubation time, but without reducing the signal from this quantitative analysis. According to some variants, the flushing is performed with the use of approximately from 50 ál to 5000 ál wash buffer solution, and mainly, approximately from 50 ál to 500 ál of wash buffer solution, in a period of time approximately from 10 to 300 seconds.

In addition, it is preferable to use several cycles of washing small amounts of wash solution, which are separated by intervals when the wash solution is not used. This sequence allows the diffusion leaching, in which the labeled antibody is diffused over time in the volume of wash solution from protected parts of the unit of analysis, such as the edges or surfaces with which these antibodies are poorly connected, then they can be removed when you wash solution is removed from the response.

In many cases, lastly the nd operation is the distribution of the enzyme's substrate for the detection conjugate using optical or electrical means. Here are examples of substrates.

For example, the reagent in the individual unit of the reagent of the claimed device may be a substrate of the enzyme for immunological analysis. In another embodiment, the transfer operation of the reagent in the form of the substrate from the individual unit of the reagent can be repeated after a reaction at the site of capture. For example, the substrate of the enzyme is transferred in place of the reaction and incubated. After measuring the received signal of the quantitative analysis used, the substrate may be removed and replaced with fresh substrate, and then measure the signal of the quantitative analysis. A signal that carries information about individual subject to detection of the analyte using the system described here, is obtained using the first and second injection of substrate. The second substrate is typically the same as the first substrate. In one embodiment, the second substrate is transferred into the location response from the second unit of the reagent of the claimed device. In another embodiment, the second substrate is transferred into the location response from the same block of the reagent, and the first substrate. Transferring the second substrate to create a second reaction, which allows to obtain a second signal representing information about the individual analyte. The intensity of the source signal and the second intensity of the second signal are compared to compute the final and tensively signal, carries information about the individual analyte, and to evaluate the accuracy of the quantitative analysis.

In one embodiment, can be used in the intensity of multiple signals for quality control of quantitative analysis. For example, if the signals differ by 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more, the results of the quantitative analysis can ignore (discard).

In one embodiment, the described method provides for re-loading of the sample and/or conjugate detection (labeled enzyme antibodies) and/or substrate of the enzyme and sample, to confirm the signal of quantitative analysis or to conduct an internal review. For example, you can re-use this tip for quantitative analysis or unit analysis to verify the functional dependence, and/or to enter additional sample or control material to obtain a second signal.

In some cases, a re-boot of the substrate in the unit of the enzyme can be carried out using the system described here, which allows you to automatically transfer the liquid sample and reagents in the units of analysis. In the case of some of the quantitative analyses do not want the system to give the results immediately or in time, so this method of testing allows you to reliably increase the TB results. The response observed after iterations supplements substrate of the enzyme, can be used to verify the source of the response or to calculate the recovery peak.

The experiments showed that by adding the second substrate of the enzyme in the unit of analysis, it is possible to maintain the reproducibility of the results. In accordance with some variations, the control method using a block analysis allows you to repeat the tests, which give the response significantly lower than expected.

In the implementation of any described here, the control means may encounter various errors (uncertainty). As examples of errors in the quantitative analysis can lead to (but without limitation) errors due to imprecise manufacturing unit of analysis or devices, due to inaccurate suction samples and/or one or more reagents, due to the inexact unit of analysis relative to the photomultiplier detection, and by skipping unit of analysis in the device or system.

In accordance with some variations of the present invention, it is proposed a method of obtaining a pharmacological data, useful for assessing efficacy and/or toxicity of drugs in laboratory animals, using published fluidic devices or systems.

When using the l is the responsibility of the animals in the preclinical evaluation of drugs, it is often necessary to kill an animal for selecting the amount of blood sufficient for conducting quantitative analysis to discover the interest of the analyte. This causes financial and ethical negative consequences, so it is preferable for the selection of such a quantity of blood to a laboratory animal didn't have to kill. In addition, it also allows you to use the same animal for testing at different points in time, which allows for more effective evaluation of the impact of medications on the same animal. On average, for example, a mouse select 6-8 ml of blood per 100 g of body weight. An advantage of the present invention is a very small volume of blood required for the implementation of preclinical trials in mice or other small laboratory animals. In accordance with some options, select average from 1 ál to 50 ál of blood. In one embodiment, taken approximately from 1 ál to 10 ál of blood. In accordance with the preferred options, select about 5 ml of blood.

Another advantage associated with the fact that laboratory animal remains alive, it becomes obvious during preclinical studies. When, for example, many mice are used to control levels of the analyte over time in the body fluids laboratory is Torno animals then the results of the test introduces a variable parameter through the use of many animals. If to control levels of the analyte over time to use only one laboratory animal, can be undertaken more accurate preclinical testing.

In accordance with some variations of the present invention, it is proposed a method of automatically monitoring the compliance of patient treatment regimens with the use of the devices or systems. The method includes the following operations: creating the conditions for reaction of the sample of bodily fluid reagent quantitative analysis device to obtain a visible signal that carries information about the presence of analyte in said sample; detecting the specified signal to the specified device; comparing the specified signal with a known profile associated with the specified treatment to determine that the patient complies with or does not comply with the treatment regimen; and informing the patient about the specified compliance or non-compliance.

In another embodiment, the system and methods in accordance with the present invention constitute the means of discovery of new biomarkers and/or means for assessing communication trends such markers with the results of treatment of diseases.

In another embodiment, the system and methods in accordance nastojasim invention make it possible to identify trends in levels of biomarkers and provide patient information on a daily adjustment of the dose to the optimum level for this patient (for example, when the adaptive range of the dose).

In accordance with some options, noncompliance with patient treatment regimens associated with taking the wrong dose of the drug, including (but without limitation) by taking multiple doses or with complete lack of dose or not proper mixing of medicines. In accordance with a preferred variant, the patient immediately report the failure to comply with treatment regimens, after comparing the signal with a known profile.

The patient or subject in a clinical trial may forget to take a sample of bodily fluid for analysis described here. In accordance with certain options, offers a way of warning the patient about the need for sampling bodily fluid using the device described here, which includes the following operations: using Protocol executable on the specified device with the specified Protocol is in an external device associated with a specified patient, and which contains the time and date of the tests specified sample of bodily fluid; and informing the patient about the need for tests specified bodily fluid at a specific date and time, if the sample has not yet been tested. In accordance with some options, the patient may be notified of the application of wireless communication, as mentioned here above. Compliance with therapeutic regimens can be improved through the use of prompts on the display and receiving responses from patients (for example, when using the touch screen).

The patient may be provided with a device when he prescribe medicine, using any known method, for example, he can get the device at the pharmacy. Similarly, passing the clinical examination subject can get these devices at the beginning of the clinical examination. Contact information about the patient or subject, including (but without limitation) the cell phone number, email address, text message address or other wireless communications devices, at this time can be entered into the external device and United, for example, with a database of patient or subject. The software on the external device may include a driver or other program which allows you to detect that the signal generated by the detection devices, has not yet been received by the external device, for example, at a specified time, and then the external device may send a warning to the patient about the need to take a sample of bodily fluid.

In one embodiment, the system directly passed to the consumer who uses her lifestyle or practice dispute the om. Suitable lifestyle, and data of physical exercises can be entered into the system, so can be carried out measurements of the parameters showing the stretching of the muscles and anaerobic metabolism (for example, about the level of lactic acid). In accordance with some variations, the system may be sufficiently low, so that it can be portable.

In another embodiment, the system is particularly well suited for measuring markers in the blood of small animals such as rats and mice, which are commonly used in preclinical trials. Such animals have a small amount of blood. So the system is a quantitative analysis requiring very small amounts of samples are particularly useful, especially when conducting long-term tests, when it is necessary to take several samples from one animal with small intervals from each other. These considerations are particularly important when it is necessary to measure in parallel multiple analytes.

In one embodiment, the system allows convenient packaging, reliable for shipment, several elements that are required to perform many complex quantitative analyses. For example, the elements for the quantitative analysis can be introduced into the housing with a snap.

Quantitative analysis

Different types of quantitative analizamos to be performed using the fluidic device in accordance with the present invention, for the detection of interest of the analyte in the sample. A wide variety of markers currently in use, can be used when carrying out the stated quantitative analyses. In accordance with some options, markers detect using spectroscopic, photochemical, biochemical, electrochemical, immunochemical or chemical means. For example, useful markers of nucleic acids include radioisotopes R, 35S, fluorescent dyes, reagents with dense electrons (electron-dense) and enzymes. In the scientific and patent literature describes a wide variety of markers suitable for labeling biological components that are generally applicable for labeling biological components in accordance with the present invention. As suitable markers can lead to radionucleotides, enzymes, substrates, cofactors, inhibitors, fluorescent components of the chemiluminescent components, bioluminescent markers or colorimetric markers. Reagents that determine the specificity of the quantitative analysis can be, for example, monoclonal antibodies, pelikanova antibodies, proteins, samples of nucleic acids or other polymers such as affinity matrix, carbohydrates or lipids. Detection can be performed with use the of any known methods, including using spectrophotometric or optical tracking radioactive, fluorescent or fluorescent markers, or other methods which track a molecule based on its size, charge or affinity. Detectable parameter can be any material that has a detectable physical or chemical property. Such detectable markers are widely used in the fields of gel electrophoresis, column chromatography, solid substrates, spectroscopic studies and so on, and, as a rule, used in areas such markers can be used in accordance with the present invention. Thus, the marker includes without limitation any song that can be detected by spectroscopic, photochemical, biochemical, immunochemical, based on the sample nucleic acid, electrical, optical and thermal or other chemicals.

According to some variants, the marker may be associated directly or indirectly with the subject detection molecule, such as a product substrate or enzyme, in accordance with well known methods. As mentioned here above, use a wide variety of markers, and the choice of marker depends on the required chustvitelnos and, ease of conjugation connections, stability requirements, available instrumentation, and opportunities for waste removal. Non-radioactive markers are often attached using indirect means. As a rule, specific for the analyte receptor is associated with generating the signal component. Sometimes receptor analyte associated with the adapter molecule (such as Biotin or avidin), with a set of reagents for the quantitative analysis contains a binder component (such as biotinylated reagent or avidin), which forms the connection with the adapter and with the analyte. The analyte is associated with a specific receptor in place of the reaction. Labeled reagent may form a complex with a sandwich structure in which the analyte is in the center. The reagent may also form a complex with the analyte for the receptor at the site of reaction or to bind free receptors at the site of reaction, not occupied by the analyte. The marker itself is detectable or may be associated with a signaling system, such as a detectable enzyme, a fluorescent compound, chemiluminescent compound or chemiluminogenic component, such as an enzyme with luminogenic substrate. Can be used several ligands and anti-ligands. When a ligand has a natural anti-ligand, for example, Biotin, thyroxine, digoxigenin and cortisol,then it can be used in combination with labeled anti-ligands. Alternatively, any heptenone or antigenome connection can be used in combination with the antibody.

According to some variants, the marker can also be anywhereman directly for connections, generating a signal, for example, by conjugation with an enzyme or fluorophore. Interest enzymes, such as markers, in the first place are hydrolases, particularly phosphatase, esterase and glycosidase, or oxireductases, particularly peroxidases. Fluorescent compounds include fluorescein and its derivatives, rhodamine and its derivatives, danceline group and umbelliferon. Chemiluminescent compounds include dioxetane, acridinium esters, luciferin and 2,3-dihydropteridine, such as luminal.

Methods of detecting markers are well known to specialists in this field. For example, when the marker is radioactive, the detection tool is the calculation of outbreaks or use the film if autoradiography. When the marker is fluorescent, it can be detected by excitation fluorochrome light of the appropriate wavelength and detecting the resulting fluorescence, for example, using microscopy, visual inspection, photographic film, and with the help of electronic means of detection, so the x as digital cameras, the charge-coupled devices (CCDs) or photomultipliers and the photocells, or other detecting devices. Similarly, enzyme markers can be detected through the use of appropriate substrates for the enzyme and detecting the corresponding reaction product. Finally, a simple colorimetric markers can often be simply detected by monitoring that is associated with the marker color. For example, gold conjugate is often pink, while various conjugate balls get the colour of the balls.

In accordance with some options, audible signal may be generated using a source of luminescence. The term luminescence is usually used to refer to radiation of light from a substance that occurs for any reason, in addition to increasing its temperature. Typically, the atoms or molecules emit photons of electromagnetic energy (e.g. light), when they move from an excited state to a lower energy (usually in the main quantum state). If the excitation creates a photon, the luminescence process is called photoluminescence. If the excitation creates an electron, the luminescence process called electroluminescence. More specifically, the electroluminescence occurs through direct insertion and removal of electrons, that is to form electron-hole pair, and subsequent recombination of electron-hole pairs from photon radiation. Luminescence, which occurs due to a chemical reaction, usually called chemiluminescence. Luminescence, which is created by living organisms, called bioluminescence. If the tube is permitted by the spin transition (e.g. transition to the singlet-singlet or triplet transition-triplet), then the process of photoluminescence can be called fluorescence. Usually fluorescent radiation disappears when removing the source of excitation, because of the transient excited States, which quickly relaxes due to the allowed spin transitions. If photoluminescence is the result prohibited by the spin transition (e.g. transition triplet-triplet), then the process of photoluminescence usually referred to as phosphorescence. Typically fluorescence emission persists for a long time after removal of the excitation source, because of the long excited States, which can recover only through such forbidden by spin transitions. Fluorescent marker can be any one of the above properties.

Suitable chemiluminescent sources contain a chemical compound, which becomes electronically excited by chemical reaction and may then emit the light, which is detected by the signal or the donor of energy to a fluorescent acceptor. Different families of chemical compounds can create a chemiluminescence under different circumstances. One of these families of chemical compounds is 2,3-dihydro-1,4-phthalazinedione. Commonly used connection is luminal, which represents the 5-amino compound. Other compounds of these collections include 5-amino-6,7,8-trimethoxy - and dimethylamine[sa]Benz similar. These connections can be made fluorescent by using alkaline hydrogen peroxide or hypochlorite of calcium and substrate. Another family of these compounds are 2,4,5-triphenylimidazole, and Latin is a common name for the original product. Chemiluminescent analogues are para-dimethylamino and methoxy substituents. Chemiluminescence can also be obtained using oxalates, and usually with the help of oxalyl active esters, for example, p-nitrophenyl and peroxide such as hydrogen peroxide under basic conditions. Other well-known useful chemiluminescent compounds are N-alkyl acridinium esters and dioxetane. Alternatively, luciferin can be used in combination with luciferase or lucigenin to get bioluminescence.

ispolzovanie herein, the term "analyte" includes (but without limitation) of the medicinal product, prodrugs, pharmaceutical drugs, drug metabolites, biomarkers such as expressed proteins and cell markers, antibodies, serum proteins, blood cholesterol and other metabolites, polysaccharides, nucleic acids, biological analytes, biomarkers, genes, proteins, or hormones, or any combination of them. The analytes can be combinations of polypeptides, glycoproteins, polysaccharides, lipids and nucleic acids.

Of particular interest are biomarkers associated with a specific disease or with a specific disease stage. Such analytes include, but are not limited to, the analytes associated with autoimmune diseases, obesity, high blood pressure, diabetes, neuronally disease and/or degenerative diseases of the muscles, heart diseases, endocrine diseases, metabolic disorders, inflammation, cardiovascular disease, sepsis, diseases of the blood vessels, with different types of cancer, Alzheimer's disease, and any combinations thereof.

Also of particular interest are biomarkers that different quantities are present in one or more tissues of the body, including the heart, liver, prostate, lung, kidney, bone marrow, blood, skin, bladder, brain, muscles, nerves, and selected the x tissues, affected by various diseases such as various cancers (malignant or not metastatic), autoimmune diseases, inflammatory or degenerative diseases.

Also of particular interest are the analytes, which are indicative of the microorganism, virus or chlamydia (chlamydiaceae). Exemplary microorganisms include, but are not limited to, bacteria, viruses, fungi and protozoa. The analytes that can be detected using the inventive method also includes the emerging blood borne pathogens selected from the group which includes (but not limited to, Staphylococcus epidermidis, Escherichia coli, resistant to methicillin Staphylococcus aureus (MSRA), Staphylococcus aureus, Staphylococcus hominis, Enterococcus faecalis, Pscudomonas aeruginosa, Staphylococcus capitis, Staphylococcus warneri, Klebsiella pneumoniae, Haemophilus influenzae, Staphylococcus simulans, Streptococcus pneumoniae and Candida albicans.

The analytes that can be detected using the inventive method, also include a variety of sexually transmitted disease selected from the group comprising: gonorrhea (Neisseria gorrhoeae), syphilis (Treponena pallidum), chlamydia (Clamyda tracomitis), not gonococcal urethritis (Ureaplasm urealyticum), yeast infection (Candida albicans), chancroid (Haemophilus ducreyi), trichomoniasis (Trichomonas vaginalis), genital herpes (HSV type I & II), HIV I, HIV II and hepatitis a, b, C, G, as well as hepatitis caused by TTV.

Additional analytes that may be identified is by using the claimed methods, include a variety of respiratory pathogens, including (but without limitation) of Pseudomonas aeruginosa that are resistant to methicillin Staphlococccus aureus (MSRA), Klebsiella pneumoniae, Haemophilis influenzae, Staphlococcus aureus, Stenotrophomonas maltophilia, Haemophilis parainfluenzae, Escherichia coli, Enterococcus faecalis, Serratia marcescens, Haemophilis parahaemolyticus, Enterococcus cloacae, Candida albicans, Moraxiella catarrhalis. Streptococcus pneumoniae, Citrobacter freundii, Enterococcus faecium, Klebsella oxytoca, Pseudomonas fluorscens, Neiseria meningitidis, Streptococcus pyogenes, Pneumocystis carinii, Klebsella pneumoniae Legionella pneumophila, Mycoplasma pneumoniae and Mycobacterium tuberculosis.

Additional exemplary markers that can be detected using the claimed methods include: theophylline, CRP, CKMB, PSA, myoglobin, SA, progesterone, Thu, 6-keto-PGF-l-alpha, and theophylline, estradiol, Latinized hormone, triglycerides, tryptase, lipoprotein cholesterol low density lipoprotein cholesterol, high density cholesterol and IGFR.

Exemplary markers of liver include (but without limitation) LDH, (LD5), (ALT), arginase 1 (like liver), alpha-fetoprotein (AFP), alkaline phosphatase, the alanine aminotransferase, the lactate dehydrogenase and bilirubin.

Exemplary markers of kidney include (but without limitation) TNFa receptor, cystatin C, urinary prostaglandin D lipocalin type, shintetsu (LPGDS), growth factor receptor of hepatocytes, polycystin 2, polycystin 1, fibronectin, uromodulin, alanine, aminopeptidase, N-acetyl-B-D-glucosaminidase, albumin and connect the surrounding retinol protein (RBP).

Exemplary markers of heart include (but without limitation) troponin I (TnI), troponin T (TnT), CK, CKMB, myoglobin, fatty acid binding protein (FABP), CRP, D-dimer, S-100 protein, BNP, NT-proBNP, PAPP-A, myeloperoxidase (MPO), the glycogen phosphorylase isoenzyme BB (GPBB), activated by thrombin inhibitor fibrinolysis (TAFI), fibrinogen, ischemia modified albumin (IMA), cardiotrophin-1, and MLC-I (light chain - I myosin).

Exemplary markers for pancreatic cancer include, but are not limited to, amylase associated with pancreatitis protein (PAP-1) and proteins regeneration (regeneratein) (REG).

Exemplary markers of muscle tissue include (but without limitation) myostatin.

Estimated blood markers include, but are not limited to, erythropoietin (EPO).

Exemplary markers of bone include, but are not limited to, crosslinked N-telopeptide of bone collagen type I (NTx), carboxyterminal cross-linking of telopeptide bone collagen, lysyl-pyridinoline (deoxypyridinoline), pyridinoline resistant titrate acid phosphatase, procollagen propecia type I, procollagen propecia type I N osteocalcin (GLA-protein, bone), alkaline phosphatase, cathepsin To, OMRS (cartilage oligomeric matrix protein), osteocrin osteoprotegerin (OPG), RANKL, sRANK, TRAP 5 (TRACP 5), a specific factor 1 of osteoblasts (OSF-1, pleiotrophin), adhesion molecules soluble glue the key, sTfR, sCD4, sCD8, sCD44 and specific factor 2 of osteoblasts (OSF-2, periostin).

In accordance with some variations of the present invention, markers are specific for the disease. Estimated cancer markers include (but without limitation) PSA (total prostate specific antigen), creatinine, acid phosphatase prostate, PSA complexes, specific gene 1, prostate, SA 12-5, the carcinoembryonic antigen (CEA), alpha Feto protein (AFP), hCG (human chorionic gonadotropin), inhibin, CAA ovarian FROM 1824, CA 27.29, CA 15-3, CAA chest 1924, Her-2, pancreatic, CA 19-9, carcinoembryonic antigen, pancreatic CAA, neuron-specific enolase, angiostatin DcR3 (soluble luring (decoy) receptor 3), endostatin, EP-HIMSELF (MK-1), free immunoglobulin light chain Kappa, free immunoglobulin light chain lambda, histatin, chromogranin And, adrenomedullin, integrin, growth factor receptor the epidermal, growth factor receptor the epidermal - tyrosinekinase, Pro-adrenomedullin N-terminal 20 peptide, vascular endothelial growth factor receptor vascular endothelial growth factor, growth factor receptor stem cells, c-kit/KDR, KDR and Midkine.

Exemplary conditions infectious diseases are (but without limitation) viremia, bacteremia and sepsis, for detection which can be used is s the following tokens: PMN elastase, complex PMN elastase/ al-PI, surfactant protein D (SP-D), HBVc antigen, HBVs antigen, anti-HBVc, anti-HIV, T-suppressor antigen cells, the ratio of antigen to T-cells, T - helper antigen cells, anti-HCV, pyrogens, P24 antigen, muramyl-dipeptide.

Exemplary markers of diabetes include, but are not limited to, C-peptide, hemoglobin Ale, picatiny albumin, the end products of advanced glycosylation (AGEs), 1,5-anhydroglucitol, gastric inhibitory polypeptide, glucose, hemoglobin, ANGPTL 3 and 4.

Exemplary markers of inflammation include, but are not limited to, rheumatoid factor (RF), antinuclear antibody (ANA), C-reactive protein (CRP), a protein of Clara cells (uteroglobin).

Exemplary markers of Allergy include (but without limitation) total IgE and specific IgE.

Exemplary markers of autism include (but without limitation) ceruloplasmin, metallothionein, zinc, copper, B6, 12, glutathione, alkaline phosphatase and activator APO-alkaline phosphatase.

Exemplary markers of clotting factor disorders include (but without limitation) b-thromboglobulin, factor 4 plaques and factor von Willebrand.

According to some variants, the marker may be specific to therapy. SOKH inhibitors include (but without limitation) THV (Mor-1), 6-keto-PGF-1-alpha (SOH 2), 11-degidro-DV-1a (Cox-1).

Friend the e markers in accordance with the present invention include, but are not limited to, leptin, the leptin receptors, and procalcitonin. Brain S100 protein. Substance P and 8-Iso-PGF-2a.

Approximate geriatric markers include (but without limitation), neuron-specific enolase, GFAP and S100B.

Exemplary markers of nutritional status include (but without limitation) prealbumin, albumin, retinol binding protein (RBP), transferrin, acylation stimulating protein (ASP), adiponectin associated with the agouti protein (AgRP), similar to angiopoietin protein 4 (ANGPTL4, FIAF), C-peptide, AFABP (protein binding adipocyte fatty acid, FABP4), EFABP (protein binding epidermal fatty acid, FABP5), glicentin, glucagon, similar to glucagon peptide-1, similar to the glucagon peptide-2, ghrelin, insulin, leptin, leptin receptor, PYY, RELMs, resistin and sTfR (receptor soluble transferrin).

Exemplary markers of lipid metabolism include, but are not limited to, various APO-lipoproteins, Apo Al, Apo-B, Apo-C-CII, Apo D, Apo-E.

Exemplary markers of the state of coagulation include (but without limitation) factor I: fibrinogen, factor II, prothrombin, factor III; tissue factor, factor IV: calcium, factor V: proaccelerin, factor VI, factor VII: proconvertin, factor VIII: anti-hemolytic factor, factor IX: Christmas factor, factor X: Stuart-Prower factor, factor XI: the thromboplastin of antecedent plasma factor XII: the factor of Hageman, factor XIII: abilities.i fibrin factor, prekallikrein, kininogen with high molecular weight, protein C, protein S, D-dimer, activator profibrinolytic fabrics, profibrinolytic, A2-doesn? t, inhibitor 1 activator profibrinolytic (RAS).

Approximate monoclonal antibodies include antibodies to EGFR, ErbB2, and igfir.

Exemplary tyrosine kinase inhibitors include, but are not limited to, Abl, Kit, DERIVED, Src, ErbB2, ErbB4, EGFR, EphB, VEGFR1-4, PDGFRb, FLt3, FGFR, PKC, Met, Tie2, RAF and TrkA.

Exemplary inhibitors of serine/ creolin kinases include (but without limitation) ACT, Aurora a/b/B, CDK, CDK (pan), CDK1-2, VEGFR2, PDGFRb, CDK4/6, MEK1-2, mTOR and RKS-beta.

GPCR targets include (but without limitation) the histamine receptors, serotonin receptors, the angiotensin receptor, adrenergic receptors, the muscarinic receptors of acetylcholine, GnRH receptors, dopamine receptors, receptors of prostaglandin and ADP receptors.

According to a separate embodiment of the present invention proposes a method of monitoring multiple pharmacological parameters useful for assessing efficacy and/or toxicity of therapeutic agents. For example, a therapeutic agent may contain any substances that have therapeutic efficacy and/or have therapeutic potential. Such substances include (but without limitation) the biological or chemical compounds, such as simple or complex organization of the organic molecules, peptides, proteins (e.g. antibodies) or polynucleotide (for example, anticavity (anti-sense)). Can be synthesized with a wide variety of compounds, including, for example, polymers, such as peptides and polynucleotides, and synthetic organic compounds based on various core structures, which can also be used as therapeutic agents. In addition, various natural sources, such as extracts of plants or animals, can give compounds for screening. It should be borne in mind that, although this is not always clearly expressed, the agent (therapeutic agent) can be used in isolation or in combination with another agent with the same or a different biological activity. In addition, proposed here, the agents and methods are also designed for combination with other therapies. It should also be borne in mind that small molecule drugs often measured by mass spectrometry, which can be inaccurate. Based on the measurement of antibodies quantitative ELISA tests are much more accurate.

Physiological parameters in accordance with the present invention include, but are not limited to, parameters such as temperature, heart rate/ pulse rate, blood pressure and respiratory rate. Pharmacodynamic parameters the factors include the concentration of biomarkers, such as proteins, nucleic acids, cells and cellular markers. Biomarkers can carry information about diseases or may be the result of actions of drugs. Pharmacokinetic parameters (PK parameters) in accordance with the present invention include (but without limitation) the concentrations of drugs and metabolites of drugs. Identification and quantification of the RK parameters in real time from a sample volume is highly desirable to assess the safety and efficacy of drugs. If the concentration of drugs and their metabolites are outside the desired range and/or if there are unforeseen metabolites due to unexpected reactions from the medication, then you should take immediate action to ensure patient safety. Similarly, if the pharmacokinetic parameters (PD) fall out of the desired range during the treatment, then you should take immediate action.

The possibility of monitoring the rate of change of the analyte concentration or PD or RK parameters during the period of time on the same object, or trend analysis of concentrations, PD or RK parameters, when they are the concentrations of drugs and their metabolites, may, at OCI to prevent potentially dangerous situations. For example, if glucose is an interest of the analyte, the concentration of glucose in a sample in a given time, as well as the rate of change of glucose concentration within a specified period of time, can be extremely useful, for example, in forecasting and exception hypoglycemic events. This trend analysis has widespread beneficial effects in the dispensing of medicines. In the case of many drugs and their metabolites, the ability to estimate the trend and the possibility of taking preventive measures is often highly desirable.

In accordance with some variations of the present invention, it is proposed commercial way of assisting the Clinician in conducting individualized treatment. Commercial method may include, after the appointment of medications, monitoring of drug therapy, to assess trends in biomarkers over time. Commercial method may include receiving at least one pharmacological parameter from the medication of the patient, and the specified operation receiving (collecting) carried out by introducing a sample of bodily fluid to react with reactants contained in the fluid device, which was passed to the specified patient and to the / establishment, which allows to obtain a visible signal, carrying the information about the specified at least one pharmacological parameter; and an appeal for comparison, using data stored in the computer memory of the medical records of the specified patient in at least one pharmacological parameter specified patient in order to help the specified Clinician in conducting individualized treatment.

Described here are devices, systems and methods allow for automatic quantification of pharmacological parameter of the patient, and an automatic comparison of the specified parameter, for example, with the medical records of the patient, which may contain the history of the monitored parameter, or with the medical records of another group of patients. Monitoring of analyte in real time by connecting an external device that can store data and to perform any type of data processing or use any algorithm allows, for example, to get a device that helps in the treatment of a typical patient and allows, for example, to compare the current patient's data with the previous data of the patient. Thus, in accordance with the present invention offers a commercial way that allows them to effectively carry out at least part of the patient monitoring in nastoyascheevremya carried out by medical personnel.

Example 1

In this example, the device, method and system in accordance with the present invention was used to perform quantitative analysis for human VEGFR2. This example demonstrates the type of quantitative analysis that can be carried out at the site of patient care. The grip surface of the unit of analysis may be covered (reagents) in the unit of analysis in accordance with the ongoing quantitative analysis, as in this example, in accordance with the quantitative analysis of VEGFR2. The inner surface of the measuring block (made of polystyrene using injection molding in accordance with the example shown in Fig.3A) was opened for consistency covering reagents by suction and pneumatic removal. The units of analysis were introduced in 20 µl of each coating reagent and incubated at room temperature for 10 minutes. In this example, used to cover the reagents in the following sequence: neutravidin (neutravidin) (20 μg/ml) in carbonate-bicarbonate buffer solution (pH 9), biotinylated "antibody capture" (monoclonal antibody introduced in VEGFR2 at a concentration of 20 μg/ml) three times buffered (triple) saline solution (pH 8), and "fixing" reagent containing 3% albumin bovine serum three times buffered saline is. After applying a sequence of coatings, the units of analysis were dried using dry air and stored dried.

Samples for analysis were then distributed (entered) in the unit of analysis in a solution of 50 mm triple buffer (pH 8) containing albumin bovine serum and isotonic sucrose for 20 minutes. Block reagent, which contains a conjugate solution of alkaline phosphatase and labeled cow intestines monoclonal antibody designed to VEGFR2 (for explicit binding epitope to the antibody on the surface of the capture) at a concentration of 250 ng/ml in stabilizing reagent (firm Biostab), was put in contact with the unit of analysis for 10 minutes. After you create the conditions for binding of the conjugate to the complex of analyte associated with the engagement surface, the unit of analysis was washed with a solution contained in the block reagent (purchased wash buffer solution firm Assay Designs). The unit of analysis was washed 5 times. Then the unit of analysis was moved to write and mixing with another reagent that is contained in another block reagent, namely, the solution of the purchase luminogenic substrate for alkaline phosphatase (KPL Phosphaglo), and incubated for 10 minutes. Then the reaction quantitative analysis the unit of analysis was found using node detection in accordance with the present invention.

is a of Fig.12 shows the response of the quantitative analysis of VEGFR2 using the above-described exemplary method. On x axis is the concentration (PG/ml) VEGFR2; and on the y-axis delayed relative luminescence (photons). The resulting graph was used for calibration of the modular unit of analysis and units of the reactants.

Example 2

Quantitative analysis for human P1GF was performed using units of analysis and units of reagents in accordance with the present invention, and the results were read (received) with the purchase of the meter. Was concurrently conducted a quantitative analysis using the same reagents using the layout described here below disposable cartridges in the layout of the reading device. The used concentration of the analyte, respectively 0,4, 80 and 400 PG/ml is Shown in Fig.13 the results of the measurements were used to calibrate the unit of analysis and unit of reagent needed for quantitative analysis P1GF person.

Example 3

Were used Magnetisation of BioMag particles with a diameter of 1.3 μm firms Bangs Laboratories, covered with material anti-rabbit IgG. The particles were dispersed at a concentration of 14 mg/ml three times buffered sucrose solution (or, alternatively, three times buffered saline) containing 3% bovine albumin serum and erythrocytes of rabbit anti-human IgG firm CedarLane, at a concentration of more than 1.15 mg/ml. Aliquots (10 ál FL the th variance) were distributed in a conical tube and dried (frozen in liquid N ₂and dried for about 24 hours at a temperature of-70C), earlier introduction into the groove in the Chuck body. Antibody rabbit (rabbit) binds as erythrocytes and covered with material anti-rabbit IgG particles, and forms a joint agglutinate particles and erythrocytes.

Freeze-dried magnetic particles were re-suspended by adding 20 ál of whole blood, and then conducted their absorption and distribution of at least 8 times (for about 1.5 min) in a conical tube.

The blood was separated due to the installation of the tip (vertical position) in a strong, horizontally oriented magnetic field. Typically received 8 ál mostly do not contain red blood cells blood plasma without the observed hemolysis of 20 μl blood samples (yield 70%). The selection of analytes (compared to blood plasma without magnetic separation) was close to 100% for protein, VEGF, P1GF, insulin, GIP and G1P-1.

Example 4

Was performed serial dilution of samples for analysis of the analyte in the system described herein. C-reactive protein (CRP) is a marker of the acute phase of the disease. Normal levels are in the range of ng/ml (the upper end of the range) to μg/ml (the lower end of the range). In any acute phase of the disease, the human liver produces CRP, and its level is in the blood can rise to hundreds of µg/ml Measurement of CRP creates problems in previously known ROS analytical systems, because of the need to measure a wide dynamic range of the analyte (the level of which varies more than 10⁵time).

Described here, the system includes a device for transferring fluid, and a cartridge or device with matrix units of analysis and units of reagents. Tips for the quantitative analysis, which have monoclonal anti-CRP associated with their inner surface, were installed in the cartridge together with diluted detection antibody (labeled alkaline phosphatase monoclonal anti-CRP (with a different epitope specificity than the tips), wash solution and substrate chemiluminogenic Alp (PhosphaGLOTM) company K. PL.

For quantitative analysis of CRP, the cartridges were introduced pre-diluted solutions of CRP, which is used without further dilution. This was followed by the processing cartridges using the system. Consistently selected CRP solution (10 μl) and antibody detection (12 ál) pipette tips, incubated for 10 min at 34°C, and then threw. The tips were washed by suction 4 times 20 ál of wash solution, until there will be no absorption of 15 µl of the substrate in the tips. After exposure for 10 min at 37°C, was measured with movoe radiation through a meter for 5 seconds. Delayed CRP concentration as a function of signal-quantitative analysis (number of photons) and the data were adjusted to 5-membered polynomial function to obtain the calibration function, as shown in Fig.14.

Example 5

Then, experiments were carried out using serial dilutions of a sample containing the analyte with a high concentration in order to get a definite response of quantitative analysis in the system described herein and the device. The CRP solutions (20 μl) were loaded into cartridges and serially diluted using the meter (up to dilutions, respectively, in 1: 50, 250, 750 and 1500 times). Diluted solutions were processed in accordance with example 4. If the concentration of the diluted CRP exceeds the calibration range of the quantitative analysis (300 ng/ml), we observed decreasing response (as shown below; these two measures).

It is shown in Fig.15 response was modeled using a modification of the binding isotherms Scatchard (S/Smax=C/(C+C0.5). Modification assumes that the response of the quantitative analysis is linearly proportional to the concentration of antibody detection, as in this example (data not shown). Any transfer of CRP in the diluted sample in the following reagent (antibody detection) will quickly react with a reagent that does not allow him to form a bond with the antigen svyazannym antibody solid phase. The reduction of the effective concentration in the proportion of transfer CRP can be accounted for by using the ratio (D-C*f)/D.

Therefore, S=Smax*(C/(C+C0.5))*(D-C*f)/D, where S is a signal of quantitative analysis, Smax represents the maximum signal (corresponding to the zero migration), represents the concentration of the analyte, C0. 5 represents the concentration of half maximal signal (without transfer), D represents the concentration of antibody detection, a f is a fractional transfer.

The values that are used to fit the data were obtained by optimizing each of the following four parameters using the technique of minimizing the root mean squared differences between data and model fit. As shown in Fig.15 was obtained an excellent fit, while values of the parameters Smax, C0.5 and D (see table 2) were close to the values that can be expected when the maximum received signal observed C0.5 and a known concentration of antibody detection. In this model, adopted by the degree of transfer is 0.034% (decimal value 3.83 E-04).

Then can be considered data in accordance with the dilution used to achieve a final concentration in each tip for quantitative analysis, and for each level of dilution were received the same response, indicating that dilution is accurate, as shown in Fig.16.

Model described here can be used to calculate the response at any given dilution and for debugging algorithms, ensuring the presence of the analyte concentration in any tip within the range of the calibration. Graphic display means of the data shown in Fig.17, and the normalized response of the quantitative analysis (In/Utah) deferred in function of the logarithm of the normalized concentration (C/C0.5) for the following relative dilutions: 1: (solid line), 5:1 (dashed line) and 25:1 (dotted line). In Fig.18 and 19 shows an example similar to the one shown in Fig.17, when other normalized concentrations. Simple pattern recognition algorithms can be used to identify reliable data at high concentration samples. For example, for most of the dose response, the signal decreases with dilution. If the signal at any dilution will be equal to or greater than the next higher dilution, the result of the analysis at this dilution should be discarded. In another example, the concentration obtained using the calibration function, shown in example 4, should have the same system error that is known dilution. If the estimated concentration for a smaller dilution less than the concentration for higher dilutions, the result of a smaller dilution should be discarded.

When the dose response quantitative analysis approaches the maximum, increases the slope of the concentration (Δ/ΔS) to the signal. In the case of quantitative analysis in which the relative signal change (ΔS/S) is mostly constant (for example, as in the system described herein), it may result in a greater change of the calculation of the concentration at higher concentrations. The suggested dilution or sledovatelno dilution allows to obtain accurate concentration achieved in immunological assays, but at much higher signal levels (e.g., more than 10 times) than for the zero signal of the analyte, but not close to the maximum signal (for example, less than 0.3 of the maximum signal). Serial dilution allows to obtain quantitative signal analysis in this range.

Due to the implementation of several estimates of the concentration of the analyte at different dilutions can be obtained mean (average) value. The average value can also be obtained by conducting parallel measurements at a single dilution. In some cases, the approach using serial dilution, which is described herein provide methods, systems and devices, allows to estimate the error due to non-linearity of dilution caused, for example, the influence of the matrix on the sample.

Example 6

The fluorescein is a well-known chemical, and also known antibodies with high affinity specific molecule. By attaching a few fractions of fluorescein to this protein, known as albumin, creates an artificial analyte that can be measured using ELISA. The example here, which is implemented on titration the microplate to show the ability of the wasp is estline such quantitative analysis, can easily be transferred to this device or system in accordance with the present invention.

Anti-fluoresceine monoclonal antibody was attached to the walls of the holes tiralongo microplate 384-hole, to create a surface capture. Quantitative analysis was carried out by adding a series of solutions to the wells and incubation at room temperature for 10 min at each stage, when you need it. Then in the wells was added 30 μl of the purchase of the drug, with known concentrations of fluorescein labeled bovine albumin (sample), with a ratio of about five fluorescein molecule. After mechanical removal of the sample, was added 30 μl of labeled alkaline phosphatase anti-fluorescein (antibody detection), with a concentration of 100 ng/ml After removal of the antibody detection, the wells were washed three times with 40 ál of wash solution ("Wash Buffer" Cat#80-1351 [firm Assay Designs, Ann Arbor, Michigan]), diluted before being used in the ratio of 1:20. Then add the substrate PhosphaGLO™ (40 μl) and read the response quantitative analysis using spectrofluorometry M5 within 0.5 sec. The response of the quantitative analysis shown in Fig.20.

Labeled with fluorescein albumin (5 μl, at various concentrations, up to 80 ng/ml) was dissolved in three times in buffered saline is aStore, which contains bovine albumin at a concentration of 3 mg/ml (buffer), was injected in polypropylene tubes and dried over night dry air. Complete drying was checked by weighing the tubes before and after drying and inspection of the corresponding loss in mass until it reaches almost a constant final weight. Analyte coated at the expense of additives 5 μl water, 20 μl of human serum and 180 ál of buffer, and then produced a stirring. Control experiments were performed by mixing 5 μl of aliquot solution of the analyte with 20 μl of serum and 180 ál of buffer.

The selection of the analyte was measured with the use described here, the quantitative analysis. As described in more detail below, the receiving signal of the quantitative analysis and the selection of the analyte) is mainly quantitative in all concentrations. It may be desirable to have a good selection (>90%) with high accuracy (<2% CV at allocation). In some cases, the dose response of the quantitative analysis can be made linear in interest in the range due to the use of low concentrations of analyte and excess reagents. For example, a linear dose response quantitative analysis can be achieved due to the significant capacity of binding antigen on the surface of the grip, so even PR is the highest level of the analyte only reasonable proportion (for example, <30%) locations (sites) will be busy at the end of the binding assays. As mentioned here above, for analytes in the range of ng/ml and quantitative analyses with small incubation times (e.g., <30 min), this condition can be ensured when described here above the floor surface capture. In accordance with another example, use a sufficient concentration of antibody detection, so that this concentration is mainly not depleted during incubation, antibody detection (for example, <30% reagent to be associated with the surface at the highest levels of the antigen), and this condition can be achieved by the use of concentrations of antibody detection, comprising approximately from 5 to 100 ng/ml In accordance with another example, a linear dose response quantitative analysis can be achieved due to the generation of a signal that is smaller than the linear characteristic of the detection devices (for example, PMT with a linear response up to 4 million photons per second). Described here are systems and methods allow you to work within this range. In accordance with another example, a linear dose response quantitative analysis can be achieved by establishing a sufficiently strong signal to be accurately measured (for example, with the speed of counting photons, orientirov is but above 1,000 photons per second).

Described here are the tips for the quantitative analysis cover by suction to the following sequence of reagents: 20 ál at a concentration of 5 μg/ml anti-fluorescein rabbit (Molecular Probes # A6413) in carbonate buffer solution pH 9, 20 μl of 3% bovine albumin in three times in buffered saline pH 8, and 20 μl at a concentration of 2.5 μg/ml bovine serum albumin labeled with fluorescein (Sigma-Aldrich A9771), and after each suction spend incubation for 10 min and removal of the fluid. The nibs are then washed three times by suction bovine albumin in three times in buffered saline pH 8, followed by incubation with 3% bovine albumin in three times in buffered saline pH 8. After that, the tips dried as described here above. These tips were used for the quantitative analysis of samples containing goat (goat) anti-fluorescein, through incubation, 20 μl of aliquot the following solutions in sequence: goat anti-fluorescein (sample) three times buffered saline, pH 8, containing 3% BSA; labeled alkaline phosphatase rabbit-anti-goat the fluorescein at a concentration of 100 ng/ml in solvent Stabilzyme™ (commercially available), with washing four times with wash solution Wash Buffer; and PhosphaGLO™ chemiluminogenic substrate of alkaline phosphatase, with the conduct of each is incubatee at room temperature for 10 minutes The quantitative analysis was performed by counting photons received within a time of about 10 seconds, the meter using a photomultiplier tube (luminometer M5 firms Molecular Devices), with the installation of each tip in a custom-built frame, which is joined to the platform tiralongo microplate. Obtained in this example the results are shown in Fig.21. In Fig.21 shows linear characteristics similar to those shown in Fig.20.

Example 7

This example shows the predictability of the response immunoassay for CRP using these tips for the quantitative analysis, when carried out the initial additive reagents, removal of the reaction product, washing tips and then re-introduction of some or all components of the quantitative analysis. Use the following sequence of operations quantitative analysis: incubation arrowheads in m the othe meters at 34°C for 10 min, sequentially: (1) breakdown (CRP 0.3,3, 30, 150 and 300 µg/ml), diluted with use of the measuring device 500, and then in 2000; (2) labeled with alkaline phosphatase rabbit anti-goat IgG ["Dab"] (5 ng/ml), rinsed three times; and (3) with PhosphaGLO™, chemiluminogenic substrate of alkaline phosphatase ["Substrate"]. The experiment was conducted on several measures, when reading the rate of appearance of photons within 10 seconds after the operation 3. The final (at the tip) CRP concentration was 0.15, 0.6, 1.5, 6, 15, 60, 75, 300 and 600 ng/ml, and levels of illumination ranged from 2,000 to 600,000 counts (photons) within 0.5 sec. In some experiments, after the operation (3) quantitative analysis, made the removal of the reaction product and carried out in various ways repeat: operations 3 (diamonds and solid line in Fig.22), operations 2+3 (squares and dashed line) or operations 1+2+3 (triangles and dashed line). The results were shown in Fig.22 as a signal to re-quantitative analysis in function of the signal source quantitative analysis.

The signals repeated quantitative analysis linearly related (proportional) to the signal source of the quantitative analysis. The second Supplement of the substrate gives a stronger signal compared to the original signal, while repeated quantitative analyses, which were introduced together Dab and substr is t, and put together a sample, Dab and the substrate, give weaker signals compared to the original signal. In accordance with one example of the use of this method, all operations in the sequence of operations quantitative analysis can be subjected to quality control, to understand that they are in accordance with the expected ratio between the first and subsequent iterations of operations quantitative analysis.

For example, as described here, if the operation of the quantitative analysis does not proceed properly, then the result of the quantitative analysis can be discarded as incorrect or the later iterations of quantitative analysis can be used as an appropriate response quantitative analysis.

An immunological assay for C-reactive protein was carried out in the system described herein. Six equivalent tips for the quantitative analysis were incubated sequentially breakdown (200 ng/ml CRP); labeled alkaline phosphatase rabbit anti-goat IgG, with flushing; and PhosphaGLO™, chemiluminogenic substrate of alkaline phosphatase. Incubation was carried out for 10 min at 34°C. the Experiment was conducted on three measures, when read (rate of occurrence) of photons within 10 seconds. On average was found about 40,000 counts (photons) in order for a read time of 0.5 sec. In this example, the level of illumination on the tips 1 and 2 meter 3 gives quite different results, as shown in the following Table 3. The meter was then used to wash the tips and for the introduction of fresh PhosphaGLO™ substrate (suction 2). The results were presented as the ratio of the degree of illumination for each tip to the average value for the six lugs on each respective meter. After the second suction nozzles 1 and 2 give results that are linearly related to the other four lugs in the meter 3, which suggests that solved the problem of low signal in tips 1 and 2.

1. The device for automatic detection of the analyte in the PR shall be a bodily fluid, which contains:
matrix addressable blocks of the analysis performed with the possibility of a chemical reaction, which gives a distinct signal that carries information about the presence or absence of the analyte;
matrix addressable blocks of reagents, which refer to individual adisoemarto block reagent matrix corresponding to the individual adisoemarto block analysis matrix units of analysis, and in which the individual unit of the reagent is performed with calibration reference signal corresponding to the individual unit of analysis, to the Assembly of the matrices on the device, with the individual unit of analysis includes the tip for the quantitative analysis, and the tip for the quantitative analysis has an inner surface with reagents, fixed on the surface for detection of the analyte.

2. The device for automatic detection of analyte in a sample of bodily fluid that contains:
block sampling made with the possibility of taking samples of bodily fluid;
the matrix blocks of the analysis performed with the receiving portion of the sample from unit sampling and executed with the possibility of a chemical reaction, which gives a distinct signal that carries information about the presence of the analyte in the sample; and
the matrix blocks reagents made with the possibility of the content is to be reagents for chemical reactions;
moreover, the individual unit of analysis matrix units of analysis includes the tip for the quantitative analysis performed with the opportunity to take the reagents, when at least one of the blocks of the analysis and at least one of the blocks of the reagent to come into fluid communication with each other, and the tip for the quantitative analysis has an inner surface with reagents, fixed on the surface for detection of the analyte.

3. The device under item 1, which further comprises a block sampling, configured to receive a sample of bodily fluid.

4. The device under item 1 or 2, in which the individual unit of the reagent is configured to receive the rolling unit of analysis.

5. The device under item 1 or 2, in which the individual unit of analysis is executed with the possibility of immunological analysis.

6. The device under item 1 or 2, in which the sample of bodily fluid is a blood sample.

7. The device under item 2 or 3, in which the unit of sampling is configured to receive a sample volume of bodily fluid about 20 μl or less.

8. The device under item 2 or 3, in which the unit of sampling is configured to receive a sample volume of bodily fluid, which represents a drop of blood.

9. The device under item 1 or 2, which further comprises the preliminary block on the processing, configured to select portions of the sample of bodily fluid for chemical reactions to detect the analyte.

10. The device under item 9, in which the sample of bodily fluid is a sample of whole blood, and a portion of the sample is a blood plasma.

11. System for automatic detection of analyte in a sample of bodily fluid, which contains:
a. the device under item 1 or 2; and
b. node discovery, designed to detect visible signal, which carries information about the presence or absence of the analyte.

12. System on p. 11, which further comprises a programmable mechanical device configured to move the individual unit of analysis within the device.

13. System on p. 11, which further comprises a device for transferring fluid.

14. The system under item 13, in which the device to transfer fluid is a pipette.

15. The system under item 13, in which the device for transferring fluid is an automatic device.

16. System on p. 11, which further comprises a communication unit for transmitting information about the analyte to be detected.

17. System on p. 11, which further comprises a heating unit configured to receive the individual unit of analysis.

18. System on p. 11, which additionally contain what it magnetic block.

19. System for automatic detection of multiple analytes in a sample of bodily fluid, which contains:
fluid device, which contains:
block sampling made with the possibility of the contents of the sample of bodily fluid;
the matrix units of analysis, and the individual unit of analysis of this matrix blocks analysis is configured for carrying out a chemical reaction, which gives a signal that carries information about the individual analyte specified set to be the detection of analytes; and
the matrix blocks, reagents, and individual block reagent specified matrix blocks contains reagents reagent;
moreover, the individual unit of analysis includes the tip for the quantitative analysis, and the tip for the quantitative analysis has an inner surface with reagents, fixed on the surface, for detecting the analyte; and
device for transferring fluid, which includes many heads, and each head is made with the possibility of entrance into engagement with the individual unit of analysis, with a device for transferring fluid includes a programmable processing device, configured to direct transfer of the sample of bodily fluid from a block of sample and reagent from the individual unit of the reagent in the individual unit of analysis is A.

20. System on p. 19, in which the configuration of the processing device for direct transfer fluid allows you to set the degree of dilution of the sample of bodily fluid in the matrix blocks of the analysis, in order to receive signals carrying the information of the plurality of analytes to be detected within the detection range.

21. The system under item 20, in which the sample of bodily fluid contains at least two analyte present at concentrations that differ by at least 2 orders of magnitude.

22. The system under item 20, in which the sample of bodily fluid contains at least two analyte present at concentrations that differ by at least 5 orders of magnitude.

23. System on p. 21, in which a given degree of dilution of the sample of bodily fluid enables the registration of signals carrying information of at least two analytes.

24. System on p. 20, which further comprises a detection unit, configured to detect the intensity of the signal in the detection range.

25. System on p. 24, in which the detecting device is a photomultiplier tube.

26. System on p. 25, in which the detection range is approximately from 20 to 10 million counts per second.

27. System on p. 19, in which the sample of bodily fluid has a volume of approximately what else than 20 μl.

28. System on p. 19, in which the sample of bodily fluid is a single drop of blood.

29. System on p. 19, in which the individual head configured for connection with the individual unit of analysis.

30. System on p. 19, in which the individual unit of analysis provides a place immunological reaction analysis.

31. System on p. 19, in which the tip of the individual unit of analysis is the tip of the pipette.

32. System on p. 19, in which the device to transfer fluid is a pipette.

33. System p. 32, in which the pipette is a pipette with the displacement of air.

34. System on p. 19, in which the device for transferring fluid further comprises a motor connected to the programmable processing device.

35. System p. 34, in which the motor moves the specified set of heads on the basis of information from the programmable processing device.

36. System for automatic detection of analyte in a portion of the plasma sample of whole blood, which contains:
a. the device according to p. 10; and
b. node discovery, designed to detect visible signal, which carries information about the presence or absence of the analyte.

37. The method for detecting analyte in a sample of bodily fluid, which includes the following operations is:
a. submission of blood samples in the device under item 1 or 2;
b. creating conditions for response specified sample within at least one unit of analysis; and
c. the detection of a specified audible signal of the presence of analyte in said sample of bodily fluid.

38. The method according to p. 37, in which the sample of bodily fluid is a blood sample, and the method further provides for the selection of plasma from blood samples.

39. The method of Assembly of the device for automatic detection of analyte in a sample of bodily fluid, comprising a housing that contains: a matrix of addressable units of analysis, in which the individual unit of analysis matrix made with the possibility of carrying out a chemical reaction, which gives a distinct signal that carries information about the presence or absence of the analyte; a matrix of addressable blocks of reagents, which apply to the individual unit of the reagent matrix corresponding to the individual unit of analysis matrix, and the method includes the following operations:
(i) placement in the body, in accordance with the subject to detect the analyte, matrix addressable units of analysis and individual analysis block matrix made with the possibility of carrying out a chemical reaction, which allows to detect interest of the analyte, and the individual unit of analysis contains Nakonechny is for the quantitative analysis, which includes an inner surface with reagents, fixed on the surface, for detecting the analyte;
(ii) placement in the body, in accordance with the subject to detect the analyte, the matrix blocks, reagents, and individual block reagent matrix reagents corresponds to the individual unit of analysis; and
(iii) the consolidation of the matrices (i) and (ii) inside the unit.

40. The method according to p. 39, which additionally provides the choice should be detected analyte.

41. The method according to p. 39, which is provided for sealing the device.

42. The method according to p. 39, which additionally provides for the marking device label, which is subject to detection of the analyte.

43. The method according to p. 42, in which the label is a barcode or RFID tag.

44. A method of automatic detection of multiple analytes in a sample of bodily fluid, which includes the following operations:
a. the introduction of the sample of bodily fluid in the fluid device, and a fluid device includes: a block of samples made with the possibility of the contents of the sample of bodily fluid; matrix units of analysis, and the individual unit of analysis of this matrix blocks of the analysis performed with the possibility of a chemical reaction, which gives a signal that carries information about an individual Ana is it specified set of detectable analytes; the matrix blocks, reagents, and individual block reagent specified matrix blocks of the reactants contains a reagent, and the individual unit of analysis includes the tip for the quantitative analysis, and the tip for the quantitative analysis has an inner surface with reagents, fixed on the surface, for detecting the analyte;
b. the connection of the individual unit of analysis to a device for transferring fluid;
c. the transfer of the sample of bodily fluid from the block sampling in the individual unit of analysis using the device to transfer fluid; and
d. transfer the reagent from the individual unit of the reagent in the individual unit of analysis, in order to carry out the reaction of the reagent with a sample of bodily fluid and to receive a signal carrying information about the individual analyte from a variety of subject detection of analytes.

45. The method according to p. 44, in which said device for transferring fluid contains a lot of heads, with each head configured to enter into engagement with the individual unit of analysis; and a programmable processing device, configured for direct transfer of the sample of bodily fluid from a block of sample and reagent from the individual unit of the reagent in the individual unit of analysis.

46. The method according to p. 45, which additionally provide the supports the introduction of teams in the programmable processing device.

47. The method according to p. 46, in which team controls the transfer operation of the sample of bodily fluid in the individual unit of analysis.

48. The method according to p. 44, in which the transfer operation of the sample of bodily fluid includes a change in the degree of dilution of the sample of bodily fluid in the individual unit of analysis, to obtain a signal that carries information about the individual analyte many to be the detection of an analyte within the detection range.

49. The method according to p. 44, in which the sample of bodily fluid contains at least two individual analyte that is present at concentrations that differ by at least 2 orders of magnitude.

50. The method according to p. 44, in which the sample of bodily fluid contains at least two individual analyte that is present at concentrations that differ by at least 5 orders of magnitude.

51. The method according to p. 49, in which the preset degree of dilution of the sample of bodily fluid enables the registration of signals carrying information of at least two individual analytes.

52. The method according to p. 44, in which the detection range is approximately from 1000 to 1 million samples per second when using a photomultiplier.

53. The method according to p. 44, in which the volume of the sample of bodily fluid is approximately less than 20 μl.

54. The method according to p. 44, W is the sample of bodily fluid is a single drop of blood.

55. The method according to p. 44, in which the reagent in the individual block reagent is a substrate of the enzyme for immunological analysis.

56. The method of selection of plasma from blood samples using the system on p. 19, which includes the following operations:
a. mixing blood samples in the presence of magnetized particles in the unit of sampling, and the magnetized particles contain surface of the capture antibody to bind not plasma portions of the blood sample; and
b. the imposition of the magnetic field over the area of collection of blood plasma, for mixing blood samples, to obtain a suspension plasma portions of the blood sample on top of the collection zone of the plasma.

57. The method according to p. 56, in which the unit of sampling is a capillary tube.

58. The method according to p. 56, in which the volume of the blood sample is approximately less than 20 μl.

59. The method according to p. 56, which take the blood plasma in the amount of approximately less than 10 μl.

60. The method according to p. 56, in which the blood sample is not diluted.

61. The method according to p. 56, in which the mixing takes place in the presence of antibody, unbound with a hard surface.

62. The method according to p. 56, in which the mixing is the mixing due to the action of the syringe.

63. Way of an automated immunoassay for the detection of analyte present in the tion of plasma samples of whole blood, which includes the following operations:
a. submission of samples of whole blood in the device according to p. 10;
b. detection signal, which carries information about the presence or absence of analyte in said sample of bodily fluid; and
c. transfer of the results of the operation (b) to the user.

64. The method according to p. 63, in which an immunological assay is an ELISA.

65. The method according to p. 63, in which the results are passed via wireless communication.