Container for isolation and identification of microorganism. RU patent 2510844.

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. Claimed is container for isolation and identification of microorganism. Container includes upper part, which has wide internal diameter, and lower part, which has capillary tube, middle conic part, connecting said upper and lower parts, and optic window on the bottom and/or on one or more than one wall of container. Optic window is less than 0.1 inch (2.54 mm) thick and is transparent for wavelength of near-infrared, visible and/or ultraviolet light spectrum. Window contains quartz, quartz glass, sapphire, acrylic resin, methacrylate, cyclic olefin copolymer, cycloolefin polymer or any their combination. Capillary tube has internal diameter from 0.01 inch (0.03 mm) to 0.04 inch (1.02 mm).

EFFECT: container ensures isolation of microorganisms, which are free of interfering materials and compatible with fast identification technologies, from hemoculture and other complex samples.

8 cl, 12 dwg, 1 ex

CROSS REFERENCE TO RELATED APPLICATION

This application claims the priority concerning the provisional application at the U.S. patent №61/110187, titled "Method and System for Detection and/or Characterization of a Biological Particle in a Sample", filed October 31, 2008, which is included in this proposal.

THE SCOPE OF THE INVENTION

The present invention is directed to the separator for the separation of microorganisms. In particular, the separator of the present invention can be used for separation of microorganisms for the characteristics and/or identification.

PRIOR ART

Detection of pathogenic microorganisms in biological fluids should be implemented in the shortest possible time, in particular, in the case of septicemia, for which the mortality rate remains high, despite a wide range of antibiotics are available to doctors. The presence of biologically active agents, such as microorganisms, body fluids of a patient, particularly in the blood, usually defined using vials hemoculture. Infection of the circulatory system are associated with high morbidity and mortality, in addition, implementation of modern methods of diagnosis, cultivation and subsequent biochemical identification and testing for sensitivity to antibiotics may take several days. Usually start empirical therapy on the basis of clinical symptoms and test results affect clinical decisions only if initial treatment fails. The ability to describe infection of blood circulation within the first few hours, preferably within hours, after the positive result of hemoculture would significantly strengthen the clinical relevance of the provided diagnostic information. To fill this demand, methods of molecular amplification, but this approach remain serious problems. The environment itself positive hemoculture is a naturally amplified the population of micro-organisms with the potential use of rapid tests identification (ID).

Traditional automatic phenotypic ID tests such as system Vitek®, Phoenix™ and Microscan®, or manual phenotypic tests such as API, require that the microorganisms were in the appropriate phase of growth and to be free from interfering environments and blood products order to obtain reliable results. These systems use colonies grown from a positive culture for 18-24 hours on Wednesday in Petri dishes. However, if you desire to get quicker results of some lab reported the use of these systems with microorganisms isolated from bottles with positive hemoculture. These tests directly from the bottle" is suitable not for all of microorganisms (e.g., unfit for gram-positive cocci), not confirmed by the manufacturers of tests and, as a rule, take 3-8 hours to produce results. Faster and more widely specific tests are urgently needed in order to ensure doctor clinically relevant results within the first few hours, preferably within hours, after the positive result of culture.

The methods of optical spectroscopy, such as your own fluorescence (IF), infrared spectroscopy (FTIR) or Raman spectroscopy, and mass spectrometry, such as MALDI-TOF (ionization time-of-flight laser desorption using the matrix), have the potential, offering the possibility of identification of microorganisms very quickly but they may encounter interference from many highly fluorescent and absorbing compounds present in the liquid microbiological culture media and in clinical specimens such as blood, or in combination. The most frequently used methods of isolation of microorganisms directly from positive hemoculture are two-stage differential centrifugation and centrifuging in a test tube to separate the serum. However, these methods have several disadvantages. The resulting product microorganisms often contains polluting erythrocytes, platelets, lipid particles, plasma enzymes and cellular debris, which can cause bad results in the traditional ID tests. These methods are also very time consuming and unsafe due stages, which can result in aerosol exposure of potentially dangerous pathogens on user. Need simple, safe and reliable methods of isolation of microorganisms from hemoculture and other complex specimens, which is free from these interfering materials and compatible with the technology fast identification.

SUMMARY OF THE INVENTION

The present invention is directed to a cage or container that can be used for separation of microorganisms from a sample that contains or is suspected on the content of microorganisms. In accordance with the present invention, the separator can be used to partition or deposition unknown microorganism and subsequent studies divided sample or sludge characteristics and/or identification of unknown microorganism.

In one aspect of the present invention is directed to a container for isolation and identification of microorganisms, including:

(a) the upper part, with wide internal diameter;

(b) the lower part with a narrow inner diameter; and

(c) optical window on the bottom, top, and/or one or more than one wall of the container, which is transparent at least for part of the near-infrared, visible and/or ultraviolet light spectrum. Optional this container can optionally have average conic section, linking a wide inner diameter of the upper part with a narrow inner diameter of the bottom.

In another aspect of the present invention is directed to one-time separator, including:

(a) the cylindrical container the form containing the body having the longitudinal axis, which defines the longitudinal internal capillary tube, oriented along the axis of having a first end and a second end, where the body is additionally defines the tank, which is connected with the first end of the capillary tube;

(b) where a building near the second the end of the capillary tube is made of optically transparent material;

(c) the cap of the tank to provide access to the tank, giving the opportunity to dispense liquid sample into the tank.

Optional cylindrical container can contain density buffer inside the tank. The container can optionally have a conic section, connecting the tank and capillary tube.

In one form of implementation of the present invention microbial agent, split or precipitate on the bottom of the capillary tube located in the cage or container in such a way as described in this proposal. Divided or besieged microbial agent, you can explore the characteristics and/or identify this microbial agent.

In another form of implementation of the separator can be sealed, for example, the device can be hermetically closed. Such devices can take advantage of the safe handling of potentially infectious agents. In other possible forms of implementation of the separator can provide the means to access separated, isolated or besieged sample of the organism, by giving it the ability to extract a sample from the separator before the test, or for additional testing.

The container can contain polypropylene ball.

SHORT DESCRIPTION OF GRAPHIC MATERIALS

Figure 1 is a promising picture of the separator in accordance with one form of implementation of the present invention.

Figure 2 is a type of cross section separator figure 1.

Figure 3 represents a promising picture of the separator in accordance with another form of the implementation of the present invention.

Figure 4 presents a view of the cross-section of the upper part of the separator, shown in figure 3.

Figure 5 presents a view of a transverse section of the lower part of the separator, shown in figure 3. The lower part of the separator fitted to the lower end of the upper part of the separator 4.

6 is a type of cross section separator figure 3 shows the split of microbial agent in the section of the capillary tube separator (e.g. sediment after centrifugation).

Fig.7 is a type of cross section of different forms of implementation bottom of the separator figure 3. As shown, this form of exercise has two opposite sides with notches, leading to an adjacent narrow lateral walls, thus giving the opportunity to explore divided microbial agent in the section of the capillary tube from the walls of the cage.

Fig is a schematic illustration of concentrated microbial agent in the separator 6, which research through the bottom of the tube by means of the module of study.

Figure 9 is a promising picture another alternative forms of implementation of the separator of the present invention.

Figure 10 is a type of cross section separator Fig.9.

11 represents a promising alternative image of the cover for a separator of the present invention.

On Fig shown a photograph of the separator in accordance with one form of implementation of the present invention. On the photo are clearly visible lysed sample density buffer and a Deposit of the microorganism in accordance with the present invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention can be implemented in various forms and should not be seen as limited by the forms of implementation, as set out in this application. Most likely, these forms of implementation submitted to this description was more thorough and complete and fully passed the volume of the invention specialists in the art. For example, signs, illustrated in respect of one form of exercise that can be included in other forms of exercise, and signs, illustrated with respect to specific forms of its implementation, can be removed from this form of implementation. In addition, various changes and amendments to the suggested forms of implementation that does not deviate from the present invention will be obvious to specialists in a given field of technology in the light of the present description.

Methods of separation, characteristics and/or identification of microorganisms revealed the following applications for U.S. patents, held by the same holder: (1) serial hall, entitled "Method for the Isolation and Identification of Microorganisms", filed October 30, 2009; (2) serial hall, entitled "Method for Separation, Characterization and/or Identification of Microorganisms using Spectroscopy", filed October 30, 2009; (3) the serial hall, entitled "Method for Separation, Characterization and/or Identification of Microorganisms using Mass Spectrometry", filed October 30, 2009; and (4) serial hall, entitled "Method for Separation, Characterization and/or Identification of Microorganisms using Raman Spectroscopy", filed October 30, 2009. These applications are included in this proposal by reference. Briefly, these authors of inventions disclosed ways isolation, characterization and/or identification of microorganisms in the sample. These methods allow the division, characteristics and/or identification of microorganisms faster than the methods of prior art that results in faster diagnoses (for example, for a subject, suffering or suspected to septicaemia) and the characterization/identification of contaminated materials (for example, food products and pharmaceuticals). In these and other ways characteristics and/or identification of microorganisms is often necessary to provide separated, isolated or deposited a sample of the microorganism for subsequent methods characteristics and/or identification. In the present invention disclosed separator that can be used for separation, isolation and/or deposition of microorganisms from the sample. For example, the separator of the present invention can be used for deposition of micro-organisms (for example, by centrifugation) of liquid culture (for example, hemoculture). The sediment of the organism may be one or more than one stage of the research for measurements that are useful for the characterization and/or identity of the organism.

In one form the implementation stage of the research can be done when separated, isolated or deposited a sample of the organism remains in the separator. For example, hermetic separator (for example, a tightly closed device) you can use to get separated, isolated or deposited a sample of the microorganism, and then this divided, isolated or deposited a sample of the microorganism can be non-invasive method of study to obtain data or measurements capable of characterization and/or identity of the organism. In another form of implementation separated, isolated or deposited a sample of the microorganism can be extracted from the separator before the examination. For example, separated, isolated or deposited microorganism can be resuspendiruetsa in the appropriate buffer and remove (for example, a pipette) from the device or container. In another form of exercise, as disclosed in this application, the cage or container may include lower part that can be removed or taken away from the cage or container (that is, a detachable bottom). When working this bottom part can be severed from the cage or container to provide access to separated or isolated micro-organisms in it.

In General, the cage or container of the present invention can be any device or container, useful for separation, isolation, or deposition of microorganisms of the test specimen, containing or suspected on the content of microorganisms. For example, the cage or container may include one - or multi-part of the body and the cap or cap. The device can be obtained by moulding, casting, blown or laminated with other well-known methods in the art. As a rule, any known plastic, glass or transparent material or the like can be used for the separator. The separator will be formed so that it had a hole at one end, giving access to the inside of the machine or container for loading and/or unloading of the tested samples. In the form of the implementation of the cage or container includes the building of cylindrical form, which is closed at one and is open at the opposite end. Typical cover or cap, you can use any known mechanism for closing or other sealing the inner parts of the unit or container from the external environment. For example, a cap or cap can be a cover with a latch that is affixed to the device, or container, and you can lock it in place over the hole of a device or container to close or seal inner part of the device or container from the external environment. Alternative cover may be a cover with thread, which can be screwed on the device or container to close the device/container. As is well known in the art, cover may have a thread on the inner sides of the cover, which are turning or screw on the thread, located on the outer wall of a device or container. In one form of the cover may contain one or more rubber 0-ring on the inner surface, as is well known in the art. The use of one or more than one 0-ring provides a seal (for example, hermetic sealing). In another possible form of implementation, as shown in figure 11, cover or cap 100 can have sharp wall 104, which can pierce, for example, needle or the like, by giving it the possibility of delivery of test samples in a tightly closed device or a container. Using sharp partitions 104 can take advantage of security for the user or lab assistant when handling potentially infectious agents and provide automation of means of identification. Peelable partition 104 also ensures that the device or container remains sealed (for example, hermetically closed), and therefore provides protection from possible contamination of the separator.

The tested samples that can be subject to separation, isolation, or settling in the cage or container of the present invention, include both clinical and non-clinical samples, where the presence and/or the growth of microorganism exist or it can be suspect, and sample materials that usually or occasionally tested for the presence of microorganisms. For example, the test sample may be a culture medium from culture clinical or non-clinical sample. Typical samples that you can cultivate, and then analyze the method of separation for separation, isolation, or deposition of microorganisms contained in them, can enable blood, serum, plasma, blood fraction of, joint fluids, urine, semen, saliva, faeces, cerebrospinal fluid, the contents of the stomach, vaginal secretion, tissue homogenates, punctate bone marrow, bone homogenates, sputum, punctate, strokes and washing water smears, other body fluids, and the like.

In one form of realization of a device or container has the upper inner chamber or reservoir, which has a wide diameter to keep the test sample and most of the density of the buffer, and lower internal camera or capillary tube having a narrow diameter for the collection is divided, isolated or deposited microorganisms. The top inside of the camera or a reservoir can have an internal diameter of about 0,32 (8,13 mm) to about 0,40 (10,16 mm) inches, for example, from about 0,34 (8,64 mm) to about 0,38 (9,65 mm) inches, for example, about 0,36 (9,14 mm) inches. For divisions in the microscale internal diameters can be even less. For example, the internal diameter of the narrowest part can range from about 0,001 (0.03 mm) to about 0,04 (1,02 mm) inches, for example, from about 0,002 (0.05 mm) to about 0,01 (0,254 mm) inches. Lower chamber or capillary tube can have an internal diameter from around 0.04 (1,02 mm) to about 0,12 (3.04 from mm) inches, for example, from about 0,06 (1.52 mm) to about 0,10 (2.54 mm) inches, for example, about 0,08 (2,032 mm) inches.

In another form of realization of a device or the container is a one-time separator, including tubular container that contains the body, which has the longitudinal axis, where the case determines the longitudinal internal capillary tube, oriented along the axis of having a first end and a second end, where this case additional specifies the tank, which is connected with the first end of the capillary tube. One aspect of this form of implementation of the hull near the other end of the capillary tube is made of optically transparent material. The tank is equipped with a detachable lid or cap, which provides access to the tank, allowing to dispense liquid sample into the tank. Optional density buffer can be pre-Packed in a device or container.

The cage or container may also have an average conical part or camera, connecting the upper inner chamber or the tank from the bottom of internal camera or a capillary tube. Inner side walls average cone can have a conical shape or may reduce the diameter between the top inside of the camera or reservoir and lower internal camera or a capillary tube. These inner lateral wall of the cone can have an angle of about 20 to about 70 degrees, for example, from about 30 to about 60 degrees. In one form of the lower, narrow part is less than half of the total height of the container, for example, less than about 40%, 30%, 20% or 10% of the total height of the container.

In some forms of implementation container designed to separated, isolated or besieged microorganisms can be easily distinguished from the container after the separation, either manually or automated way (so that the technicians were not affected by the contents of the container). For example, the container can contain detachable part or the detachable part, which contains the sediment, and which can be separated from the rest part of the container. In another form of implementation, the container provides a means of access to the precipitate after the separation, such as one or more than one hole or one or more than one permeable surface to insert the syringe or another dosing or to remove sediment. In one form of the container can be a test tube, such as the centrifuge tube. In another form of implementation of the container can be a chip or card. In one form of the container is a standalone container, that is, a device for separation of one sample.

The container can contain optical window through which you can carry out the study. Optical window can be on the bottom, top, and/or on the walls of the container. This window can be made of any material that is transparent to light (for example, at least part of the near infrared (NIR; 700 nm - 1400 nm)ultraviolet (UV; 190 nm - 400 nm) and/or visible (view, 400 nm to 700 nm) light spectrum. Examples of suitable materials include, without limitation acrylic resin, acrylic, quartz, quartz glass, sapphire, cyclic olefin copolymer (COC) and/or cycloolefines polymer (SOR) (for example, Zeonex® (Zeonex®, San Diego, CA). In one form of implementation container made entirely from a material optical window. In another form of implementation of the container can be made (for example, moulded) of two or more than two separate parts, such as optical UV-visible-NIR transparent component for optical Windows and other material (for example, the standard molded plastic low cost) for making the rest of the container. In one form of the optical window is fine enough to allow spectroscopic study, which will depend on the material of the window. In another form of implementation is fine as far as possible, to minimize interference with spectroscopic study. For example, the window may have a thickness of less than about 0,20 (5,08 mm) inches, for example, less than approximately 0.15 (3,8 mm), 0,10 (2.54 mm) or 0.05 (1.27 mm) inches.

Now with reference to graphic materials the following illustrates some of the possible configurations of the cage or container of the present invention. One possible form of implementation of the separator is shown in Fig.1-2. As shown in figure 1 and 2, the separator 2 includes the lower part 6, which usually has a cylindrical shape, and the upper part is defined outside ribbed structure or the tab 8, 9 hole and shut the hood 4. The lower part 6 includes building 10 container, which holds the inner chamber, which includes the upper reservoir 14, the average conical section 16 and the bottom of the capillary tube 18, where they are arranged around the longitudinal axis of the container. As shown, the average cone section 16 connects the upper reservoir 14 wider inner diameter and capillary tube 18 smaller diameter. As a rule, building 10 container can be molded or shaped into different way from any known plastics material, known in the art. Coming out rib structure or ledge 8 can function as a stop valve cap 4 and/or may provide grounds, enabling improved capture device by the user. The upper part of the device can also contain threaded 12 on the outer side of the unit 2 for screwing or screwing the valve cap 4 on the device, 2, through the closing or sealing the inner chamber. The device may additionally contain a thin optical window 19, through which you can carry out the study. In one form of implementation of the diameter of the optical window 19 can be designed to fit fiber optic cable and contribute to the precise device is connected to the spectrometer. As described above, optical window includes 19 section of the device, which consists of material, which is transparent to light, and through which you can carry out the study. In other forms of carrying out the device can be completely made from a material that is transparent to light, by allowing it to conduct research.

In some forms of implementation of the separator 2 of this form of exercise can be pre-loaded density buffer 43 (shown, for example, figure 6-7)to facilitate the separation, isolation, or deposition of microorganisms. In another form of the density buffer can be added to the separator 2 immediately before the sample application that you want to expose phase separation is described in this proposal. In one other form of implementation of the separator 2 can be pre-loaded density buffer 43 and the solution for lysis (not shown)to facilitate the lysis of the sample and separation, isolation, or deposition of microorganisms, as described in patent applications in the US, held by the same holder, are referenced in this application.

Another form of the lower sections of the separator is shown in Fig.7. The lower section 48 can be movably attached or fixed to the upper section 22 with the formation of the separator 39 in accordance with this invention. The top section 22 and the lower section 47 include the upper body is 32 and lower case 49, respectively, that define the internal chamber. Internal camera includes the upper reservoir (not shown), the average conical section 42 and capillary tube is 45. As shown, the bottom case 34 additionally contains the outer walls are sloped inward 48, which resulted in a thin lateral walls on the opposite sides of the bottom of the inner capillary tube 45. These thin side walls enable research separated, isolated or deposited microorganism 50 through the wall of the cage.

One other form of implementation of the separator 60 shown in figure 9-10. Referring to these figures, the separator consists of 60 corps 62, which defines the inner tank 80, average conical section 82 and bottom of the capillary tube 84. Average cone section 82 connects the upper reservoir 80 larger diameter from the bottom of the capillary tube 84. The upper reservoir 80 has access through removable lid or cap 72, which can screw or screw on the thread 66, created on the upper outer wall of the body of 62. In accordance with this form of the bottom part of the housing 62 contains four fixing wing 68 used to ensure the sustainability of the separator 60 vertical standing, such as a Desk. In another form of exercise four grooves on the bottom of the wings 68 create a deep area for the exact connection of fiber-optic cable. Centering excitation beam in this way results in improved reproducibility of fluorescence and reduced contamination of the signal emitted randomly scattered light. Separator 60 additionally contains optical box 70, formed in the body of 62 at the bottom of the capillary tube 84. Optical box 70 includes a small section of the reduced thickness on the housing 62, through which you can explore separated, isolated or deposited microorganism. As described in this application, optical window 70 can be made of optically transparent material.

In some forms of implementation separator 60 of this form of exercise can be pre-loaded density buffer 85 (as shown, for example, figure 10)to facilitate the separation, isolation, or deposition of microorganisms. In another form of the density buffer can be added to the separator 60 immediately before the sample application that you want to expose phase separation is described in this proposal. Yet in any other form of exercise separator 60 can be pre-loaded density buffer and the solution for lysis in order to facilitate the lysis of the sample and separation, isolation, or deposition of microorganisms.

On Fig shown operation research concentrated microbial agent within 50 separator 60. In one form of implementation separated, isolated or deposited microorganisms can be explored by any known means in this field equipment (listed in the application as a means of research 58). For examples, as disclosed in jointly filed a patent application in the US, serial hall, entitled "Method for the Isolation and Identification of Microorganisms", filed October 30, 2009, stage of the research can be carried out using own fluorescence spectroscopy, Raman spectroscopy or other optical method.

Although the above form implementation concentrated microbial agent explore when he is inside the cage, also consider that separated, isolated or deposited a sample of the microorganism can be removed from the separator and investigated, for example, using mass spectrometry, as disclosed in jointly filed a patent application in the US, serial hall, entitled "Method for Separation, Characterization and/or Identification of Microorganisms using Mass Spectrometry", filed October 30, 2009.

As noted in the application above, phase separation, isolation, or precipitation can be done to share microorganisms from other components of the sample (for example, micro-organisms or their components), and concentration of microorganisms in a divided, isolated or besieged sample, which can be researched in order to identify and characteristics. The separation stage or deposition does not have to be complete, i.e. there is no need to happen 100% of the division. It is only necessary that the division of microorganisms from other components of the sample was sufficient to give the possibility to study microorganisms without significant interference from the other components. For example, the division can result in collapse of microorganisms, which is clean at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 96, 97, 98 or 99% or higher.

In one form of exercise, as described more fully in applications for U.S. patents, held by the same holder, discussed in this application, the division shall be implemented by stages centrifugation at which the sample (for example, lysed the sample) are placed on top of density buffer in the separation container, and the container centrifuged in conditions which allows you to isolate the micro-organisms (e.g. micro-organisms can form the sediment on the bottom and/or on the walls of the container). In accordance with this form of exercise other components of the sample (e.g., organisms or their components that may be present in the environment of the sample) remain on top of density buffer or inside the upper part of the density of the buffer. This separation stage isolates of microorganisms from materials in the sample, such as the environment, cellular debris and/or other components that might interfere with the study of microorganisms (for example, due to the intrinsic fluorescence). In one form of the density buffer also serves to separate living organisms from dead organisms (that do not pass through density buffer). In another form of the density buffer does not contain the density gradient, either before or after centrifugation. In other words, separating the container does not centrifuged within a reasonable amount of time and/or with sufficient acceleration of material forming density buffer to create a density gradient.

The density of the selected buffer so that the microorganisms in the sample is passed through this buffer, while other components of the sample (for example, cultural environment hemoculture, cell debris) remained on top of density buffer or not passed all the way through density buffer. Density buffer can also be selected so that to share live microorganisms (which pass through the gradient) and the dead microorganisms (which do not pass through the buffer. Suitable density will depend on the material used in density buffer, and from the sample, subject to division. In one form of the density of the buffer is in the range of approximately 1,025 to approximately 1,120 g/ml, for example, from approximately 1,030 to approximately 1,070 g/ml, approximately 1,040 to approximately 1,060 g/ml or at any interval from approximately 1,025 to approximately 1,120 g/ml In another form of the density is about buffer 1,025, 1,030, 1,035, 1,040, 1,045, 1,050, 1,055, 1,060, 1,065, 1,070, 1,075, 1,080, 1,085, 1,090, 1,095, 1,100, 1,105, 1,110, 1,115 or 1,120 g/ml

Material density buffer can be any material that has the appropriate range of density for ways on this invention. In one form the implementation of this material is a colloidal silica. Colloidal silica can be without coverage (for example, Ludox® (W.R.Grace, CT)) or coated, for example, silane (for example, PureSperm® (Nidacon Int'l, Sweden) or Isolate® (Irvine Scientific, Santa Ana, CA)) or polyvinylpyrrolidone (for example, Percoll, Percoll Plus (Sigma-Aldrich, St. Louis, MO)). In one form of implementation of the selected colloidal silica, manifesting the least interference with spectroscopic study, for example, a material with the lowest native fluorescence. Colloidal silica can be diluted in any suitable environment with the formation of the appropriate density, for example, in a balanced salt solution, physiological solution and/or 0.25 M sucrose. The corresponding densities can be obtained with colloidal silica in concentration from about 15% to about 80% V/V, for example, from about 20% to about 65%.about. Other suitable material for the density of buffers is iodinated contrast agent, for example, iohexol (Omnipaque™, NycoPrep™ or Nycodenz®) and iodixanol (Visipaque™ or OptiPrep software. The corresponding densities can be obtained by iohexol or iodixanol in concentration from about 10% to about 25% wt./about., for example, from approximately 14% to approximately 18% wt./about., for samples of hemoculture. Sucrose can be used as the density of the buffer in concentration from about 10% to about 30% wt./about., for example, from about 15% to about 20% wt./about., for samples of hemoculture. Other suitable materials that can be used to obtain the density of the buffer include oil low viscosity, high density, such as immersion oil for microscope (for example, Type DF; Cargille Labs, new York), mineral oil (for example, Drakeol®5, Draketex 50, Peneteck®; Penreco Co., Pennsylvania), silicone oil (polydimethylsiloxane), Fluorosilicone oil, silicone gel, metrizoate-Ficoll® (LymphoPrep™), for example, in concentration from around 75% to about 100% for samples of hemoculture, diatrizoat-dextran (PolymorphoPrep™), for example, in concentration from approximately 25% to around 50% for samples of hemoculture, carboxymethyl cellulose, hypromellose, polyethylene oxide (high molecular weight), Pluronic® F127, Pluronic® F68, mixtures of compounds Pluronic®, polyacrylic acid, made of polyvinyl alcohol, stitched polivinilpirrolidon, copolymer PEG methyl ester and methacrylate, pectin, agar, xanthan gum, 'gellan, Phytagel®, sorbitol, Ficoll® (for example, Ficoll® 400 in concentration from about 10% to 15% for samples of hemoculture), glycerin, dextran (for example, in concentration from about 10% to 15% for samples hemoculture), glycogen, caesium chloride (for example, in concentration from about 15% to about 25% for samples of hemoculture), perfluorocarbon fluids (for example, PERFLUORO-n-octane), gidrogeologiya fluid (for example, Vertrel XF) and the like, which are well known in the art. In one form of the density of the selected buffer of one or more than one of colloidal silica, iodixanol, yogexola, cesium chloride, metrizoate-Picolla®, diatrizoat-dextran, sucrose, Picolla® 400 and/or dextran in any combination. Density buffer may consist of a combination of materials, for example a combination of colloidal silica and oil. Some combinations of the above compounds can be useful for stages of separation and read the present invention, for example combination of compounds with different properties damping UV light, such as caesium chloride and iohexol.

Once separated, isolated or deposited a sample of the microorganism is received, you can implement the next stage of the research to get the measurements that are useful for the characterization and/or identity of the organism. Useful tools of study known in the art. Additional funds research described in the applications for U.S. patents, held by the same holder, discussed in this application above.

EXAMPLES

EXAMPLE 1. Devices and methods for identification cleaned in situ sediment organisms

To explore the potential of rapid partitioning and in situ identification of microorganisms in the separator, a number of devices have been developed and made of plastic, transparent to UV light. These devices had a few common features, including the cover, reservoir for sample and conical bottom area of the optical quality to ensure spectroscopic investigations sedimentating sludge microorganisms below and/or on the wall, and indicators that facilitate the device is connected to a spectrophotometer. These devices must be able to withstand relatively high g-forces during stage separation. It was constructed a few iterations of this test tubes to improve the selection of microorganisms, reproducibility fluorescence, and to reduce contamination randomly scattered light. The tube was also designed as a tightly closed.

Optical study sedimentating sludge microorganisms was achieved either by inserting a separator in a custom adapter that is placed inside the office for sample spectrophotometer or by direct connection separator with an extensive a six conductor 300-400-micron fiber-optic cable (Ocean Optics, Dunedin, FL)joined spectrophotometer (Fluorolog® 3 (HORIBA Jobin Yvon Inc., New Jersey)). Trehseriynyy fiber optic adapter was designed to ensure the use of both systems detectors (FEG and CCD). Spectra full matrix of excitation-emission (EEM) filmed on each Deposit of microorganisms (range scan: agitation 260-800 nm; emission 260-1100 nm; increments of 5 nm).

Research reproducibility and reliability calibration was performed on disposable device configuration and fiber optic cable using a solution of purified tryptophan and Riboflavin. Target CV<2.5% were obtained for both fluorophores, which confirms the quality disposable devices and research platform.

It is proved that these devices can be used to split and study of microorganisms from the culture medium. On Fig shows an example of a device after separation by centrifuging lysed sample hemoculture containing S.aureus, using the density of the buffer. On the photo are clearly visible lysed sample density buffer and a Deposit of microorganisms in accordance with the present invention.

1. Container for isolation and identification of microorganisms, including: (a) the upper part, with wide internal diameter; (b) the lower part with capillary a tube with an inside diameter from 0.001 inch (0.03 mm) up to 0.04 in (1,02 mm); (c) the average conical part that connects these upper and lower part; and (d) the optical window at the bottom or on one or more than one wall of the container of a thickness of less than 0.1 inches (2.54 mm), which is transparent to wavelengths near-infrared, visible and/or ultraviolet light spectrum, and where the specified window contains quartz, quartz glass, sapphire, acrylic, methacrylate copolymer of cyclic olefin polymer of cycloolefine or any combination of them.

2. Container according to claim 1, which has a volume of 0.1 ml to 25 ml

3. Container according to claim 1, where the lower part is less than half of the total height of the container.

4. Container according to claim 1, which includes the device for closing or has a thread for connecting the device for closing so that the container can be sealed.

5. Container according to claim 1, which further comprises a solution with a uniform density, where this solution is selected from the group consisting of colloidal silica, iodized contrast agent, sucrose, immersion oils for microscopes, mineral oils, silicone oils, oil film, silicone gel, diatrizoat-dextran, carboxymethylcellulose, gidroksipropilmetilzelluloza, of polyethylene oxide high molecular weight, polyoxyalkylene ether, polyacrylic acid, made of polyvinyl alcohol made of polivinilpirrolidon, copolymer PEG methyl ester and methacrylate, pectin, agarose, xanthan gum, 'gellan, Gellan gum, sorbitan copolymer of sucrose and epichlorohydrin, glycerin, dextran, glycogen, cesium chloride, perfluorocarbon fluids, hydrotherapeutic liquids and their combinations.

6. Container according to claim 5, which additionally contains a selective solution for lysis.

7. Container 6, which additionally includes the thin membrane separating solution with a uniform density and the solution for lysis.

8. Container according to claim 5, which additionally includes polypropylene ball.