Apparatus and method of controlling temperature of reaction mixture


FIELD: chemistry.

SUBSTANCE: apparatus for controlling temperature of a reaction mixture contained in a reaction vessel comprises: an infrared source for exposing the reaction vessel to radiation in order to heat the reaction mixture, a temperature sensor for measuring temperature, which is an indicator of temperature of the reaction mixture and a controller for controlling the radiation source in accordance with temperature of the reaction mixture in order to selectively heat the reaction mixture. The method of controlling temperature of the reaction mixture involves determining temperature of the reaction mixture using information obtained from the temperature sensor, controlling the radiation source which is designed to expose the reaction vessel to radiation with subsequent heating of the reaction mixture, wherein the radiation source is controlled by the controller in accordance with temperature of the reaction mixture, as a result of which said temperature is controlled.

EFFECT: improved control of temperature of reaction mixtures, enabling real-time analysis of a reaction taking place in a vessel and with sufficiently high efficiency.

20 cl, 12 dwg

 

The technical FIELD

The present invention relates to devices and methods for controlling temperature of the reaction mixture and in particular to devices of cyclic heat treatment for nucleic acid amplification. However, it should be understood that the invention is not limited to the above scope.

BACKGROUND of INVENTION

References in this description to any prior publication, or on information that is derived from such publications, or any known sources are not and should not be construed as a confirmation or recognition, or as some form of indication that such publication of such information or such sources form part of the General knowledge in the field to which the present invention relates.

Polymerase chain reaction (PCR) is a technology that includes a repeating cycles that provide exponential increase in the number of polynucleotide sequences whenever you execute one of these cycles. PCR is widely known and described in many publications, including "PCR: a practical approach", .J.McPherson and others, IRL Press (1991), "PCR Protocols: a guide to methods and applications, Innis et al. Academic Press (1990), and "PCR Technology: principles and application of the program for DNA amplification", .A.Eriich, Stockton Press (1989). PCR is also described in many U.S. patents, including 4,683,195; 4,683,202; 4,800,159; 4,965,188; 4,889,818; 5,075,216; 5,079,352; 5,104,792; 5,023,171; 5,091,310 and 5,066,584.

PCR typically includes a step denaturirovannyj of polynucleotide, after which perform stage of annealing (denaturirovannyj) at least two oligonucleotides (primers) to denatured polynucleotide, namely, the hybridization of the primary material to the denatured polynucleotide matrix. After the stage of denaturirovannyj enzyme with polymerase activity catalyzes the synthesis of a new polynucleotide chain, which includes oligonucleotide (primer) and uses the original denatured polynucleotide as matrix synthesis. These stages, denaturing, denaturirovannaya primers and extension (elongation) of the primer, make up the PCR cycle.

As of repetitions, the number of newly synthesized polynucleotide increases exponentially as the newly synthesized polynucleotide from the previous cycle can act as a matrix for the synthesis in subsequent cycles. Primary oligonucleotides usually choose pairs that can denaturiruet in the opposite chain of the specified double-stranded polynucleotide sequence, so that the area between two renaturiruemykh areas increases.

Denator the tion of DNA typically occurs at a temperature of about 90-95°, resaturate primers to denatured DNA is typically carried out at temperatures of 40-60°C, and elongation denaturirovannykh primers with polymerase is usually carried out at a temperature of 70-75°C. Therefore, in the cycle of PCR, the temperature of the reaction mixture should be changed many times during execution of cyclic PCR.

PCR is widely used in many biological applications, including, for example, the analysis of DNA sequences, cloning nucleic acids sequences, site-directed mutagenesis, detection of genetic mutations, the diagnosis of viral infections, molecular "fingerprints" and control of contaminant micro-organisms in biological fluids and other sources.

In addition to PCR-known and are also used for other processes amplification in vitro, including ligase chain reaction, as described, for example, in patent US 4,988,617 issued by Landegren and Hood. It should be noted that several important processes known in biotechnology, such as hybridization of nucleic acids and sequencing, carried out under conditions of controlled temperature change solutions containing molecules of the samples. Traditional technologies based on the use of individual wells or pipes passing through zones with different temperatures. For example, in the technique known p is lichnye device cyclic heat treatment, used in amplification and sequencing of DNA, in which the reaction mixture is held unit with adjustable temperature, which varies in time. The advantage of such devices is the ability to simultaneously handle a relatively large number of samples, for example, are widely used tablets with 96 wells. However, such devices are characterized by various disadvantages, for example, their cycles of temperature change is sufficiently long, they consume a lot of power, temperature control is far from ideal and the definition of the reaction mixture in situ is difficult.

To resolve these shortcomings have been developed various devices cyclic heat treatment, in which the containers from the reaction mixtures are held in a rotating carousel shop, established inside the chamber, which provides heating and cooling. One such device is described, for example, in patent US 7,081,226 issued Wittwer, etc. However, such devices also have various disadvantages. For example, the regulation of the temperature of the reaction mixtures is far from perfect, the speed control heating and cooling of reaction mixtures also not perfect, and such devices have a relatively low efficiency.

Thus, there is a need for devices cyclic thermobreak the PCR, providing improved temperature control of the reaction mixtures, which have a fairly simple design, can provide real-time analysis of the reaction vessels with samples, and have a fairly high efficiency.

In the present invention attempts to remedy at least one of the disadvantages of the known devices or reduce their effect or to provide a useful alternative.

The INVENTION

In the present invention proposes a device for controlling the temperature of the reaction mixture contained within the reaction vessel, which contains:

a) a radiation source for applying radiation to the reaction vessel, the resulting reaction mixture is heated;

b) a temperature sensor for measuring temperature, which is the temperature of the reaction mixture; and

c) a controller for controlling the radiation source in accordance with the temperature of the reaction mixture for sample heating.

In most embodiments of the invention the device comprises a heat source for heating the chamber containing the reaction vessel.

In most cases the controller provides:

(a) raising the temperature of the reaction mixture at least partially using a radiation source and

b) maintaining the temperature of the reaction mixture at least partially using a radiation source.

In most variants the device comprises a cooling means for cooling the reaction mixture.

In most cases the coolant provides cooling of the reaction mixture at an elevated temperature.

In most cases the cooling means supplies the external air into the chamber containing the reaction vessel.

In most variants coolant serve chilled the fluid in the chamber containing the reaction vessel.

In most variants as a temperature sensor using an infrared sensor.

In most variants as a temperature sensor uses an optical sensor to measure the color of the additive-indicator in the reaction mixture, temperature-sensitive.

In most cases the temperature sensor measures the temperature of the reaction mixture.

In most cases the sensor measures the temperature of the reaction vessel, and the controller ensures the determination of the temperature of the reaction mixture using the temperature of the reaction vessel.

In most cases the sensor measures the temperature of the chamber, and the controller provides for the determination of the temperature of the reaction mixture used is of the temperature of the chamber.

In most cases, the radiation source generates infrared radiation.

In most cases, the radiation source generates optical radiation.

In most variants the device comprises a chamber for accommodating therein the used reaction vessels.

In most variants the device includes mounting fixtures for the installation of reaction vessels, and the radiation source and mountings are arranged in such a manner that a heating of the one or more reaction vessels.

In most variants the device includes a drive for moving the mounting of the armature relative to the radiation source.

In most cases, the controller provides the drive control for selectively heating the reaction mixture in the respective reaction vessels.

In most cases, the radiation source generates a zone of heating radiation, and the controller controls the heating of the reaction mixture by selective pressure of the heating zone to the reaction vessel.

In most cases the controller is a system of information processing.

In most cases the controller provides:

(a) raising the temperature of the reaction mixture to a first temperature for denaturation of polynucleotides in the reaction mixture;

b) raising the temperature of the tours of the reaction mixture to a second temperature for renaturation (annealing) polynucleotides in the reaction mixture; and

c) raising the temperature of the reaction mixture to a third temperature for hybridization denaturirovannykh of polynucleotides.

In most cases the controller provides:

a) determining the temperature of the reaction mixture using information obtained from the temperature sensor; and

b) controlling the radiation source in accordance with the temperature of the reaction mixture, which may control this temperature.

In most cases the controller provides:

a) controlling the radiation source to raise the temperature of the reaction mixture to a first temperature;

b) controlling the radiation source to maintain the first temperature of the reaction mixture;

c) controlling a cooling means for lowering the temperature of the reaction mixture to a second temperature and maintaining the second temperature; and

d) controlling the radiation source to raise the temperature of the reaction mixture to a third temperature; and

e) controlling the radiation source for maintaining the third temperature of the reaction mixture.

In most cases the radiation source is designed for selective formation of a given heating zone, and the device comprises a channel of the cooling medium for the selective formation of the given cooling zone, procesadora heating zone and a given cooling zone formed by being near the heater and channel of the cooling medium, respectively, so that control of the temperature of the reaction mixture may be carried out by the selective pressure on the reaction vessel area heating and/or cooling zone.

In most cases the reaction vessel at least partially block the image.

In most cases, the radiation wavelength is selected in accordance with characteristics of the reaction vessel and/or characteristics of the reaction mixture.

In the present invention proposes a method of controlling the temperature of the reaction mixture contained in a reaction vessel, including implementation of the controller:

a) determining the temperature of the reaction mixture using information obtained from the temperature sensor; and

b) control the source of radiation intended for the effects of radiation on the reaction vessel, resulting in a heated reaction mixture; and a radiation source control in accordance with the temperature of the reaction mixture, which may control this temperature.

In the present invention it is also proposed a device to control the temperature of the reaction mixture contained within the reaction vessel, which contains: (i) the heater is designed for selective formation of a given heating zone, and Kahn is l of the cooling medium for the selective formation of the given cooling zone, moreover, the desired heating zone and a given cooling zone formed by being near the heater and channel of the cooling medium, respectively, so that control of the temperature of the reaction mixture may be carried out by the selective pressure on the reaction vessel area heating and/or cooling zone.

In most cases in the heater uses one or more IR emitters.

In most cases the channel of the cooling medium contains several holes located near the heater, and as a cooling medium is used, the external air.

In most cases the reaction vessels used in the form of an ordered set.

In most cases the temperature of the reaction mixture can be adjusted by the selective pressure on the reaction vessel, a heating zone or a cooling zone in accordance with a specified temperature profile.

In most variants the predetermined temperature profile is intended for the amplification of nucleic acids.

In most variants zone heating and cooling is essentially the same.

In the present invention a method of controlling the temperature of the reaction mixture contained in a reaction vessel, including:

i) ensure the unit's electric is La, designed for selective formation of a given heating zone; and

ii) providing a channel of the cooling medium, designed for the selective formation of the given cooling zone;

iii) and the desired heating zone and a given cooling zone formed by being near the heater and channel of the cooling medium, respectively; and

iv) controlling the temperature of the reaction mixture by selective pressure on the reaction vessel area heating and/or cooling zone.

In the present invention a method of controlling the temperature of the reaction mixture contained in a reaction vessel, including:

i) a selective effect on the reaction vessel of a given heating zone and/or a given cooling zone, and the desired heating zone and a given cooling zone formed by being near the heater and channel of the cooling medium, respectively.

Of the following detailed description it will be clear that various forms of the invention can be used separately or together in a variety of applications, including (without limitation) the amplification of nucleic acids.

BRIEF DESCRIPTION of DRAWINGS

The preferred embodiment of the invention is described below as an example with reference to p is alagaesia drawings, showing:

figure 1 - block diagram of one embodiment of a device for controlling temperature of the reaction mixture;

figure 2 - block diagram of one embodiment of the control algorithm temperature of the reaction mixture in the device represented in figure 1;

figure 3A is a schematic side view of the second variant of the device control the temperature of the reaction mixture;

figure 3B is a schematic view in plan of the device of figure 3A;

figure 4 - block diagram of one embodiment of the controller;

figure 5 is a perspective view from above of the third variant of the device control the temperature of the reaction mixture, which shows a rotating carousel shop, installed on vessels with reaction mixtures above the infrared heater and above the cooling channels;

figure 6 is a perspective view from above of a rotating carousel store and IR heater, shown in figure 5;

figure 7 is a top view in perspective of one embodiment the support base with the node of the IR heater/reflector and cooling channels;

figure 8 is an enlarged view of part of figure 7, showing the non-contact temperature sensor, located near the site of the infrared heater/reflector;

figure 9 - view of the device shown in figure 7, installed in the case;

figure 10 is a view similar to the view of the Figo is e 8, also visible vessels from the reaction mixture;

figure 11 is a view similar to the view of figure 5;

figure 12 is a block diagram of one embodiment of a device for controlling temperature of the reaction mixture, which shows the components of the device.

DETAILED description of the INVENTION

In the description reference is made to the accompanying drawings, in which identical reference numbers refer to identical parts.

One of the variants of the device for controlling temperature of the reaction mixture contained within the reaction vessel, is described below with reference to figure 1.

In this embodiment, the device 100 includes a chamber 101 containing the source 110 of radiation used to heat the reaction vessel 121, which contains the reaction mixture 120. The radiation source can be any source, but typically is used infrared heater that generates infrared radiation. However, in other embodiments can be used lasers, light emitting diodes or similar devices, generating an optical or infrared radiation. Radiation can be used for heating the reaction vessel, which, in turn, heats the reaction mixture.

In other embodiments, the radiation can heat one or more components in the reaction mixture directly, for example, if reaction the e vessels at least partially transmit the radiation. In this regard, it is clear that the wavelength can be selected in accordance with characteristics of the reaction vessel and/or characteristics of the reaction mixture. Thus, the characteristics of the vessel, such as, for example, the thickness of the vessel and the material used, and the characteristics of the reaction mixture, such as, for example, the composition of the mixture, can be used to select the wavelength, so that at least part of the radiation passed through the material of the reaction vessel and was absorbed by the reaction mixture. However, it should be understood that in other embodiments, on the contrary, the characteristics of the reaction vessel and/or characteristics of the reaction mixture can be selected depending on the wavelength of the radiation source.

The reaction vessels can be an ordered set, which is driven by the mechanism for moving the vessel relative to the radiation source, which may provide a selective and/or periodic exposure to radiation. This scheme will facilitate the management process of the reaction, and thus can provide simultaneous processing of multiple reaction mixtures.

Inside the chamber 101 is set sensor 130 temperature for temperature measurement, which is the temperature of the reaction mixture. The measurement temperature is URS can be performed in any suitable way, including using an infrared sensor, such as a thermoelectric Converter. In other embodiments, the reaction mixture may contain an indicator, such as, for example, signal a color additive, the color of which varies with temperature, and in this case, the temperature of the reaction mixture can be measured by the optical sensor. While the temperature of the reaction mixture can be measured directly, there are other ways to measure the temperature of the reaction vessel 121. Can be also measured the temperature inside the chamber 101, and may be determined using an algorithm, the temperature of the reaction mixture.

With sensor 130 temperature and source 110 radiation is connected to the controller 140. When the device controller 140 determines the temperature of the reaction mixture, using the information obtained from the sensor 130 temperature. The controller 140 controls the source 110 radiation in accordance with the temperature of the reaction mixture, which may control this temperature. Thus, the controller 140 can control the cyclic heat treatment of the reaction mixture, for example, for nucleic acid amplification, such as PCR.

Thus, the controller 140 to the control information, received from the sensor 130 temperature, and controls the source 110 radiation. Respectively, may be any suitable type of controller, such as the information processing system programmed accordingly, a programmable gate array or similar device.

In one variation of the heating chamber 101 can be used an additional source of heat, such as, for example, a convection heater 150, which contributes to improving and/or maintaining the temperature of the reaction mixture. Usually the work of a convection heater 150 is controlled by the controller 140 in accordance with the temperature of the reaction mixture or the temperature of the chamber 101.

In one embodiment, may also be cooled using a cooling device 160. This may be the external air, and depending on the variant embodiment of the invention can directly cool the reaction vessel or chamber 101. The operation of the cooling unit is usually controlled by the controller 140 in accordance with the temperature of the reaction mixture or the temperature in the chamber to increase the rate of cooling that occurs during temperature control.

In one embodiment, the use of a radiation source, has a direct impact on the reaction vessels for the x heating or direct heating of the reaction mixture eliminates the necessity of heating the entire chamber 101. In this case decreases the time required to heat the reaction mixture, resulting in reduced cycle time and heat treatment, respectively, the time performing PCR or other amplification processes. In this case also reduces the amount of energy required to achieve the temperature of the reaction mixture used in the implementation of such processes, and thus reduce the power consumption of the device.

In some embodiments, heating the chamber 101 can be used an additional source of heat, such as, for example, a convection heater 150, which promotes thermal stability of the reaction mixture. This can be reduced the time to reach the required temperature of the reaction mixture, and at the same time increases the temperature stability of the reaction mixture.

The use of the cooling device 160 may also contribute to further reductions in the cycle of heat treatment.

In one embodiment, may also directly measured the temperature of the reaction vessel or directly to the reaction mixture. In this case, increasing the accuracy of measuring the temperature of the reaction mixture in comparison with when the measured temperature of the air in the chamber. The result is improved accuracy of temperature control of the reaction mixture that in turn, helps to achieve maximum efficiency of the amplification process, and at the same time eliminates the need to use expensive computational algorithms for obtaining the temperature of the reaction mixture at the temperature of the air in the chamber.

The example control loop heat treatment is discussed below with reference to figure 2.

In this example, at the stage 200, the controller 140 includes a source 110 of radiation and controls the temperature of the reaction mixture, using the sensor 130 temperature. At stage 210 is determined by the achievement of the first temperature of the reaction mixture, which is typically in the range from 90°C to 95°C, and if this temperature is reached, the heating is prolonged (stage 200).

As soon as you reach the first temperature, the controller 140 controls the heating process so as to maintain the reaction mixture at the first temperature for the required first interval of time, for example, for 20-30 seconds to ensure denaturation of DNA (stage 220). It should be noted that for the first cycle hot start PCR reactions can be used a longer time intervals of the order of 1-9 minutes. The time interval can be programmed depending on the performed PCR reactions or can be defined by the optical sensor, which responds to in icator reaction mixture.

The required temperature of the reaction mixture can be maintained using any suitable technical means. Thus, in one embodiment, the controller 140 may adjust the amount of radiation generated by the source 110. Additionally it can be used a source 150 of heat, such as, for example, a convection heater.

After the completion of the denaturation temperature of the reaction mixture is reduced to a second temperature, which is typically in the range from 40°C. to 60°C. the cooling Process usually is that the controller 140 turns off the source 110 radiation and/or convection heater 150 (stage 230), resulting in the reaction mixture begins to cool, and the controller 140 controls the temperature of the reaction mixture using a sensor 130 temperature. To speed up the cooling process may also be used in the cooling unit 160. At stage 240 is determined to achieve a second temperature of the reaction mixture, and if the second temperature is not reached, the process of cooling (stage 230).

Once achieved the second temperature (stage 250), the controller 140 controls the source 110 of the radiation in such a way as to maintain the reaction mixture at the second temperature for the required second time interval, for example, within 20-40 seconds, to ensure the Oia renaturation of DNA to the primer. Similarly, the temperature of the reaction mixture can be maintained at the required level using any appropriate technical means, and the time interval can be programmed or can be defined by certain characteristics.

After that, the reaction mixture is heated to the third temperature, and the controller 140 includes the source 110 radiation and controls the temperature of the reaction mixture using a sensor 130 temperature (stage 260). On stage 270 is determined by the achievement of the third temperature of the reaction mixture, which is typically in the range from 70°C to 75°C, and if this temperature is reached, the heating is prolonged (stage 260). After reaching the third temperature (stage 280), the controller 140 maintains the temperature of the reaction mixture during the third time interval, which is the elongation of the DNA. The third time interval depends on several factors, such as, for example, used DNA polymerase, and again can be programmed or determined on the basis of certain characteristics.

The above-described example of a single cycle, and in practice for the implementation of PCR or other amplification process is used a certain number of cycles, and final stage of storage.

The following describes one variant of the device control the temperature the Oh reactions with reference to figure 3.

In this embodiment, the device 300 includes a housing 310 and the cover 312 that form the chamber 311. Luggage 311 includes a mounting valve 320 to install the carousel store 321. Carousel shop 321 contains many slots 322 to install them in reaction vessels 323 containing the reaction mixture.

Mounting the valve 320 is connected with the shaft 330, which is installed on the support 331 rotatably. With the shaft 331 connected to the drive motor 332, for example, by means of the belt 324, resulting in rotation of the carousel store 321 inside the chamber 311. In the chamber 311 has a wall 313, which separates the drive motor 332 and support 331 from carousel shop 321. Wall 313 has a hole with mesh 314, through which can pass the air stream.

Luggage 311 contains a radiation source in the form of infrared heater 340, which is usually installed on the wall 313. In one embodiment, the heater 340 contains a tray 341 and the conductor 342. When the device current passing through the conductor 342, heats the conductor, resulting infrared radiation, which is emitted by the surface of the conductor 342. Tray 341 reflects the radiation, so that it is directed to the reaction vessels 323.

In this embodiment, the wall 313 is installed optical sensor 350 to determine the status of the reaction color is ndicator the reaction mixture. The optical sensor 350 may include a light source such as a laser, and the corresponding optical sensing element for measuring reflected light.

As shown in figure 3, due to the optical sensor in one embodiment, the IR heater 340 may be only part of the perimeter of the carousel store 321, leaving a gap to provide line-of-sight between the optical sensor 350 and reaction vessels 323. However, this characteristic is not essential, and can be used in another position of the optical sensor 350, as indicated by reference number 360, so that the heater 340 may pass around the perimeter of the carousel store 321.

If the heater 340 is only on part of the perimeter of the carousel store 321, it gives some advantages. For example, in this case, by heating only a portion of the perimeter of the carousel store 321, which makes it possible to heat the reaction vessels only in a partial rotation of the shop 321, which may improve temperature stability. However, in other embodiments, more uniform heating can be obtained by using the heater, which runs around the perimeter of the carousel store 321.

In one embodiment, the optical sensor 350 acts as a temperature sensor by the color distribution of the additive-indicator, sensitive to the temperature of the reaction mixture. In one embodiment, the indicator is sensitive to temperature, can be introduced into the reaction vessel, for example, by using temperature-sensitive material in the reaction mixture or introduced into the material of the reaction vessel. It must be borne in mind that the use of an optical sensor for measuring the temperature of the reaction mixture or the reaction vessel eliminates the need to use additional sensor. This simplifies the design and cost of the entire device.

In another embodiment, can be used an additional temperature sensor, as shown by reference number 360. It can be an infrared sensor, which is installed to measure the temperature of the reaction mixture or the reaction vessel, and thus does not respond to radiation of the infrared heater 340.

In another embodiment, may be used the corresponding sensor (not shown), which measures the temperature of the air in the chamber. However, such temperature measurement does not have such sensitivity and accuracy, as a direct measurement of the temperature of the reaction vessel or reaction mixture that may reduce the effectiveness of the temperature control.

The chamber 311 is equipped with a fan 371 to circulate external air in the chamber 311. In one of the options, the ants can be used as the source 372 heat to heat the outside air before it enters the chamber 311, in the result of which will be provided by convection heating of the reaction chamber.

It should be noted that the device also typically includes a controller, a variant of which will be described below with reference to figure 4.

In this example, the controller 400 includes a processor 410, a storage device 411, the device 412 I/o (keyboard and display) and the interface 413, connected to each other via a bus 414. Interface 413 may be used to connect the controller 400 with devices such as a heater 340, the drive motor 332, the sensors 350 and 360, 371 fan and source 372 heat. The interface may also include an external interface that is used to provide connection to external peripheral devices such as barcode scanner, computer system or other similar device. Accordingly, it is necessary to understand that as the controller 400 may be any suitable data processing system, programmable gate matrix or other similar device.

During operation, the processor 410 is typically executes commands, such as software stored in a storage device 411, which determines the cyclic process of heat treatment to be implemented. This can be done by accessing the specified profiles termoobrezan the key, recorded in a storage device 411, and/or by executing commands entered using the input device.

Then the processor 410 generates control signals to control operation of the heater 340, a drive motor 332 and an additional fan 371 or source 372 heat to start the process of cyclic heat treatment. When carrying out this process, the processor 410 receives signals from one or more sensors 350, 360 and uses them to determine the temperature of the reaction mixture, and for the interpretation of these signals is used, the information recorded in the storage device 411. The processor 410 may also determine the state of the reaction, for example, using signals received from the optical sensor 350.

The processor 410 uses the temperature of the reaction mixture and additional information about the status of the response as feedback information to control the operation of the heater 340, a drive motor 332 and advanced 371 fan or a heat source 372, which results in the process of cyclic heat treatment mainly as it was described with reference to figure 2.

Another variant of the device described below with reference to figures 5 to 12, which illustrates the device 1, which is provided by control of the temperature of the reaction mixture is for amplification of nucleic acids.

Rotating the shop 2 provides a support for the reaction vessel 3 containing the reaction mixture (not shown). The reaction vessel 3 is preferably made of plastic and provide a relatively rapid equalization of temperature and measurement of the reaction mixture. The reaction vessel 3 can contain any of the reaction mixture, however, in the proposed variants of the reaction mixture used for nucleic acid amplification, and the device 1 cyclic heat treatment configured appropriately, namely, the process of cyclic heat treatment is specifically designed for the amplification of nucleic acids in accordance with the above-described profile of cyclic heat treatment.

Use at least one heater 4 to supply heat to the reaction vessels 3, and at least one channel 5 is used for supplying to the reaction vessels 3 cooling medium. The heater 4 and the channel 5 of the cooling medium to provide selective formation of a given heating zone and a given cooling zone, respectively. These zones are formed by being next to the heater 4 and channel 5 of the cooling medium, respectively, so that the temperature of the reaction mixture can be adjusted by selectively influence on the reaction vessels 3 heating zone and/or area is hladiny. Formed "specified areas" can be defined as a relatively small or limited area of space that is heated/cooled. Therefore, the introduction of the reaction vessel 3 in these areas or their impact on the reaction vessels 3 leads mainly to the heating/cooling of the reaction vessels, and not to the heating/cooling of the entire cell (not shown)in which the device 1.

The device 1 can provide a more rapid cycles of heat treatment in comparison with the known devices, resulting in reduced run time amplification. In addition to reducing cycles of heat treatment can also be improved degree of control the reaction temperature in comparison with the known devices, because heated and cooled only reaction mixture. Quality process additionally improved by measuring the actual temperature of the reaction mixture in real time and providing a feedback circuit for regulating the amount of heat provided by the heater 4, and the amount of cooling medium supplied to the reaction vessels by channel 5. Other process improvement related to the dimension of the real course of the reaction occurring in the reaction vessel 3, and using the obtained information as a control signal for driven the I amount of heat and the quantity of the cooling medium, served to the reaction vessels 3.

In the preferred embodiment is used, the heater 4 non-contact activities, such as infrared heater/emitter 6, which is placed in the lower part of the rotating carousel store 2 and in close proximity to the rotating reaction vessels 3. Infrared heater 6 preferably is a stainless steel tube having an external diameter of about 2 mm and an internal diameter of about 1.5 mm IR-heater 6 preferably has a ring shape, and the diameter of the ring is approximately equal to the diameter of the rotating carousel store 2. It should be noted that the infrared heater 6 shall be capable of delivering heat to the reaction vessels 3 so that was heated exclusively local area around the reaction vessel 3. Also preferably uses a parabolic reflector 7. The reflector 7 is designed to focus heat from the infrared heater 6, mainly on the reaction vessels 3.

Channel 5 of the cooling medium may be an annular gap adjacent to the reflector 7. However, in other embodiments, the channel 5 of the cooling medium contains many located around the circumference of the holes 8 located near the reflector 7. Holes 8 of the cooling medium is preferably arranged so the m way to the cooling medium fed directly to the reaction vessel 3. In this case, around the reaction vessel 3 is formed local cooling zone. Preferably as a cooling medium is used, the external air, but this air can be pre-cooled.

The temperature of the reaction vessel 3 can be measured during cyclic heat treatment, preferably using a thermoelectric sensor 9. The measured temperature of the reaction vessel 3 can act in a feedback loop with proportional-integral-differential regulation implemented in the control microprocessor 10, which can regulate the amount of heat or the amount of cooling medium supplied to the reaction vessels 3. It should be noted that in the process of cyclic heat treatment can be measured not only the temperature of the reaction vessel 3, but can also be controlled in the course of the reaction occurring in the reaction vessel 3. Such control may be accomplished by any means, but the preferred embodiment uses a fluorescent probe introduced into the reaction mixture.

The control is carried out preferably using a light source 11, a filter 12 and tube 13 of the photomultiplier. The progress of the reaction can also register in order to find the controlling microprocessor 10. It should be noted that the progress of the reactions occurring in the reaction vessel 3 may be used as a control signal for increasing or decreasing the temperature of the reaction vessel to increase or decrease the intensity of the reactions in the reaction vessels 3.

Below will be described the additional characteristics that can be used in the above described embodiments.

In one embodiment, the temperature of the reaction mixture can be adjusted in accordance with a specified temperature profile. This allows you to use the reaction mixture for nucleic acid amplification, and the predetermined temperature profile is intended to provide such amplification. The temperature profile can be pre-recorded in the controller or in a storage device and can be selected from a set of profiles in accordance with commands received from the input device. In another embodiment, the profile may be manually entered using the input device.

In one embodiment, the reaction vessels are combined in a kit, such as a rotating carousel store. Each reaction vessel may contain the same reaction mixture or different the reaction mixture, so that at the same time can be handled by a different reaction mixture.

The heater is usually used which is one or more IR emitters, and the channel of the cooling medium contains many holes, located next to the IR emitters. In one embodiment, the heater uses infrared emitter that produces infrared energy that is absorbed by the reaction vessels and their contents, with the result that they are heated. In such embodiments, the heating zone and the cooling is essentially the same.

In one embodiment, the "target area" is provided by heat or cooling medium in a relatively small or limited space. This is a contrast to the known devices, in which the heating/cooling of the whole chamber, inside which are the reaction vessels. By focusing or concentration of heat/cooling medium within a given local area in space in which can be placed in the reaction vessel, is provided by heating and/or cooling of the reaction vessels and their contents. In some embodiments, heated/cooled only the upper end of the reaction vessel by introducing only the upper end of the reaction vessel in these areas, and in other embodiments may heat up/cool down the lower part of the reaction vessel.

However, it should be understood that the technical means of heat, in the form of infrared heater/radiator, and technical means of cooling,in the form of a channel of the cooling medium, can be arranged in such a way that they can be heated/cooled, the entire reaction vessel without substantial heating/cooling of the whole chamber surrounding the reaction vessel. Although some heating/cooling chamber can occur. However, in the proposed device is minimized useless heating/cooling chamber due to the fact that heating and cooling occurs only in the local area around the reaction vessel.

Heating and cooling of the reaction mixture or reaction vessels, or parts thereof, instead of the heating and cooling of the entire cell containing the reaction vessels, as is done in many known devices, a number of advantages. For example, the proposed technology can provide faster cycles of heating/cooling in comparison with the known devices, in which heats/cools the whole camera. It is clear that the more rapid cycles of heat treatment of the reaction mixtures can reduce the run time of amplification.

In addition, with the direct heating and/or cooling the reaction mixture to increase the effectiveness of regulation of the reaction temperature in comparison with the known devices, because heated and cooled only reaction mixture or the reaction vessel. Also can be quickly measured the actual temperature of the reaction mixture or actiongo vessel, used in the control loop with feedback. This is one of the differences from the known devices, in which the chamber is filled with a heating or cooling fluid medium, and the actual temperature of the reaction mixture is not used as the feedback parameter.

The device can also provide precise temperature control of the reaction mixtures subjected to cyclic heat treatment in the reaction vessels. This represents a significant progress in comparison with the known devices, which can provide only relatively coarse temperature control in comparable cycles of heat treatment as usual control circuit in known devices is actually open, which regulates only the air temperature or the temperature of the entire device; and the actual temperature of the reaction mixture is not used as the main parameter of the feedback loop.

In addition, it may be more efficient use of energy, because when using the invention the number of useless heat and the cooling medium is minimal. Also can be used technical means of heating and cooling is relatively smaller in comparison with the known devices, because there is no need to load the ve and the cooling of the entire camera, decreasing the cost of manufacture of the device.

The present invention has other advantages. For example, the camera body, in which is placed a rotating carousel shop, does not require isolation or isolation uses the minimum amount of insulating material, because the loss of heat/cooling medium is minimal, additionally, you can abandon the fan used to circulate the heated/cooled air around the reaction vessel and through the camera, if you use the cooling channels.

Proposed in the present invention, the device is directed to devices cyclic heat treatment intended for nucleic acid amplification, in which the reaction vessels are kept on a circular carousel shop, established inside the chamber for rotation. The most preferred devices cyclic heat treatment for use with the device proposed in the present invention, is a family of devices Rotor-Gene™, manufactured and sold by Corbett Life Sciences Pty Limited (www.corbettlifescience.com). Other similar devices are described in PCT publications WO 92/20778 and WO 98/49340. However, it should be borne in mind that other commercially available devices cyclic heat treatment which may be modified for so they worked, in accordance with the above method.

Rotation of the reaction vessels may provide various advantages. One of the main advantages is that it ensures the possibility of controlling the process of amplification in situ. Since the rotating carousel store usually has a ring shape, the heater and the channel of the cooling medium also have an annular shape, so that the reaction vessels during rotation there is a permanent flow of heat or cooling medium. In this case, the rotation of the carousel store means for heating/cooling the reaction vessels do not need to be positioned in a particular zone heating/cooling.

In some embodiments, the channel of the cooling medium may be removed from the heater radially inward or radially outward. It should be borne in mind that the heater (or the channel of the cooling medium) can be in one or more sectors of a circle, so that the reaction vessels are subjected to intermittent heating/cooling in the course of their rotation. However, in other embodiments, the heater and the channel of the cooling medium may be located in alternate sectors of the circle, resulting in the formation of alternating zones of heating/cooling.

In one embodiment, to heat the reaction mixture can be used is Atisa non-contact heater. For example, a suitable heat source may be a microwave emitter, or in preferred embodiments, use of the infrared heater. In the case of using the infrared heater is its capacity is preferably not less than 100 watts. In one embodiment, the IR heater preferably is a stainless steel tube having an external diameter of about 2 mm and an internal diameter of about 1.5 mm In another embodiment, the infrared heater is a nichrome element wound spiral around the tube.

The infrared heater can be placed at the bottom of the camera body rotating carousel store in close proximity to the rotating reaction vessels. In one embodiment, the infrared heater is under the reaction vessels so that they overlap the top IR heater. However, in other embodiments, the infrared heater can be shifted radially outward (or inward) with respect to the reaction vessels, and in this case it is arranged so that the infrared energy radiates radially inward (or outward) in the direction of the reaction vessels, placed on a rotating carousel store.

Regardless of the placement of the heater may be arranged so that heat was supplied to the reaction vessels or reaction mixture with the creation of small local heating zone around the reaction vessels. In one embodiment, the tube is made of stainless steel mounted on ceramic insulators, which are attached to the reflector plate, and the whole design gives the direction of the received IR energy mainly on the reaction vessels.

In other embodiments, the reflector plate is arranged in such a way as to focus the heat from the infrared heater, mainly in reaction vessels. In such embodiments, the reflector plate has a curved shape in cross section, preferably the shape of a parabola. Although the preferred embodiments use a reflector, but this element is not essential for the invention.

In one embodiment, the channel of the cooling medium is a circular slot located near the site of the reflector plate and the infrared heater. However, in other embodiments, the channel of the cooling medium contains many located around the circumference of the holes located near the site of the reflector plate and the infrared heater. The channels of the cooling medium can be arranged in such a way that the cooling medium fed directly to the reaction vessels. In this case, around the reaction vessel is formed a given cooling zone.

In one embodiment, as the cooling medium is used, the external air. But this is about may be any known in the art cooling medium. Supplied to the cooling air can be pre-cooled. It should be noted that the air can be cooled by any method, for example, by first passing a flow of air along the cold side of thermoelectric unit (Peltier effect). However, in some preferred embodiments, the cooling medium is cooled in the adiabatic expansion, as is well known in the art. For example, the channel of the cooling medium can be connected with a source of compressed air, and in this case uses one or more spray nozzles.

The reaction vessels are arranged in such a way that there was a rapid equalization of temperature and ensure measurement of the reaction mixture, and in this case they can be made of glass or plastic. In one embodiment, used reaction vessels, similar to Eppendorf tubes™. The reaction vessels can contain any of the reaction mixture, however, in the proposed variants of the reaction mixture used for nucleic acid amplification, and the device cyclic heat treatment configured appropriately, namely, the process of cyclic heat treatment is specifically designed for the amplification of nucleic acids, as already described.

In one embodiment, the reaction with the UDA at least partially transmit the radiation, so the reaction mixture is at least partially exposed to the radiation, that is, there is direct heating. However, in other embodiments, the reaction vessels can absorb radiation, so that they heat up and transfer heat contained in the reaction mixture.

In one embodiment, the process of cyclic heat treatment is measured the temperature of the reaction vessel. This can be used by any sensors known in the art, but are preferably used temperature sensors contactless type. For example, can be used thermoelectric sensors and similar technical means. Through the use of suitable reaction vessels, which provide a rapid equalization of temperature, the temperature of the reaction mixture will be the same as the temperature on the surface of the reaction vessel. Therefore, after reaching the set temperature, there is no need for temperature equalization. In addition, the time temperature compensation no longer depends on the ratio of surface area to volume of the reaction vessel. Because infrared energy is focused on the reaction mixture, the intensity of heating is proportional to the power of the infrared heater and does not depend on the geometry of the tube unlike other systems cyclic heat treatment, heat conductivity is adnych or convection.

In one embodiment, the temperature of the reaction mixture measured directly, for example, when the reaction vessel passes the measured radiation, as is the case when the optical determination of the color of the indicator in the reaction mixture.

Also keep in mind that when the local heating and cooling of the reaction mixture after heating to 95°C at least part of the reaction mixture will evaporate and condense on the cold parts of the reaction vessel, which is not exposed to infrared radiation. To address this shortcoming in the process of cyclic heat treatment, the rotor rotates at high speed, to the reaction mixture, which can evaporate when heated, fell down. Another way to circumvent this is to cover the reaction mixture with oil or wax, which act as a barrier to evaporation.

It should be noted that the heater that supplies heat to the reaction vessel, and a conduit supplying to the reaction vessel cooling fluid, can be activated sequentially or simultaneously, as is well known in the art. For example, when the serial enable control of the temperature can be considered a regulation "on/off", and while enabling control of the temperature can be considered "proportional the grave" regulation. In the latter case, to control the temperature of the reaction vessel can be used by the controller proportional-integral-differential control.

In one of the embodiments of the invention a method of controlling the temperature of the reaction mixture includes: providing a heater intended for selective formation of a given heating zone; and providing a channel of the cooling medium for the selective formation of the given cooling zone; and the desired heating zone and a given cooling zone formed by being near the heater and channel of the cooling medium, respectively; and the control of the temperature of the reaction mixture by selective pressure on the reaction vessel area heating and/or cooling zone.

In another embodiment of the invention a method of controlling the temperature of the reaction mixture includes: selective effects on the reaction vessel of a given heating zone and/or a given cooling zone, and the desired heating zone and a given cooling zone formed by being near the heater and channel of the cooling medium, respectively.

In these variants is provided by heating/cooling the reaction vessel without heating/cooling of the entire cell containing the reaction vessels, as it assests who is in known devices. This reduces the amount of energy needed for heating and cooling the reaction mixture, as well as reduced heating time, as indicated above.

If the description is not expressly indicated otherwise, throughout the description and in the claims the word "contains" and the like should be understood as "includes, but is not as "contains only", that is, must be understood in the sense of "including without limitation".

Except of existing options or when otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used in the description, should be understood in all cases, as indicated with the word "approximately".

Notwithstanding that the numerical ranges and parameters setting advanced scope of the invention, are approximate values, the numerical values shown in the specific examples, are given as accurately as possible. However, any numerical value principally contains certain errors that inevitably arise as a result of standard deviations obtained in the performance of their measurements.

Terminology used in this description is intended only to describe specific options device control temperature of the reaction mixtures and in no way limits the scope of the invention. If not specified in the e, all technical and scientific terms used in this description have the meanings common among specialists in this field of technology. The numeric ranges boundary values includes all numbers included in this range (e.g. the range includes 1-5 1; 1,5; 2, 2,75; 3; 3,80; 4; 5 etc).

The terms "preferred" and "preferably" indicate that under certain conditions can be obtained some advantages. However, when the same or other conditions preferred can be other options. In addition, an indication of one or more preferred embodiments of the invention does not mean that can not be used and other options, that is, other options are not excluded from the scope of the invention.

Signs of different options can be used together or interchangeably, and all the options considered in the present description, are just a few examples.

Although the invention has been described with reference to specific options, you must understand that experts in the art can implement it in many other forms. In particular, the signs of any of these options can be used in any combination in any other described embodiments.

1. The control device temperature of the reaction mixture, which finds the I inside the reaction vessel, contains:
a) a radiation source for applying radiation to the reaction vessel, the resulting reaction mixture is heated;
b) infrared temperature sensor for temperature measurement, which is the temperature of the reaction mixture; and
C) a controller for controlling the radiation source in accordance with the temperature of the reaction mixture for the purpose of its selective heating.

2. The device according to claim 1, which contains a heat source for heating the chamber containing the reaction vessel.

3. The device according to claim 2, in which the controller provides:
a) raising the temperature of the reaction mixture at least partially using a radiation source; and
b) maintaining the temperature of the reaction mixture at least partially using a radiation source.

4. The device according to claim 1, additionally containing a cooling means for cooling the reaction mixture.

5. The device according to claim 4, in which the coolant is to cool the reaction mixture at an elevated temperature.

6. The device according to claim 4 or 5, in which the cooling means supplies the external air into the chamber containing the reaction vessel.

7. The device according to claim 4 or 5, in which the cooling means serve chilled the fluid in the chamber containing the reaction vessel.

8. The device according to the .1, in which the temperature sensor measures the temperature of the chamber and the controller ensures the determination of the temperature of the reaction mixture at the temperature of the chamber.

9. The device according to claim 1, wherein the radiation source generates infrared radiation.

10. The device according to claim 1 containing chamber to contain the used reaction vessels.

11. The device according to claim 1, containing mounting fixtures for the installation of reaction vessels, and the radiation source and mountings are arranged in such a manner that a heating of the one or more reaction vessels.

12. The device according to claim 11, containing the actuator to move the mounting of the armature relative to the radiation source.

13. The device according to item 12, in which the controller provides the drive control for selectively heating the reaction mixture in the respective reaction vessels.

14. The device according to claim 1, wherein the radiation source generates a zone of heating radiation, and the controller controls the heating of the reaction mixture by selective pressure on the reaction vessel heating area.

15. The device according to claim 1, in which the controller uses the information processing system.

16. The device according to claim 1, in which the controller provides:
a) determining the temperature of the reaction mixture using the information received is coming from the temperature sensor; and
b) controlling the radiation source in accordance with the temperature of the reaction mixture, which may control this temperature.

17. The device according to claim 1, in which the heater is designed for selective formation of a given heating zone, and the device comprises a channel of the cooling medium for the selective formation of the given cooling zone; and the desired heating zone and a given cooling zone is formed near the heater and channel of the cooling medium, respectively, so that the temperature of the reaction mixture can be controlled by the selective pressure on the reaction vessel area heating and/or cooling zone.

18. The device according to claim 1, in which the reaction vessel is at least partially permeable to the radiation.

19. The device according to p, in which the wavelength is selected in accordance with characteristics of the reaction vessel and/or characteristics of the reaction mixture.

20. The method of controlling the temperature of the reaction mixture contained in a reaction vessel, including:
a) determining the temperature of the reaction mixture using the information received from the temperature sensor; and
b) controlling the source of radiation intended for the effects of radiation on the reaction vessel, resulting in a heated reaction is mesh; moreover, the radiation source is controlled in accordance with the temperature of the reaction mixture, which results in the management of this temperature.



 

Same patents:

FIELD: medicine.

SUBSTANCE: invention can find application for the purpose of prediction of recurrence-free survival in cervical cancer (CC). The method involves patient's catalase activity and malondialdehyde in peripheral blood erythrocytes at the initial CC. If erythrocyte catalase activity is within 2.2 to 9.5 mmole/min/l, and if malondialdehyde is within 215.8 to 397.1 mcmole/l, the probability of 18-month recurrence-free survival makes 60%, and erythrocyte catalase activity within 10.5 to 19.4 mmole/min/l, the probability of 18-month recurrence-free survival makes 86%.

EFFECT: use of the presented invention provides the prediction of the recurrence-free survival period in local cervical cancer.

FIELD: biotechnology.

SUBSTANCE: invention relates to determining the activity of pyruvate dehydrogenase complex (PDH complex) by 13C-MP detection (magnetic resonance detection based on the isotope 13C). The essence of the method consists in that the change in activity of PDH complex in the subject to be examined by 13C-MR detection (magnetic resonance detection based on the isotope 13C) is determined using the medium of visualisation comprising hyperpolarised 13C-pyruvate, and when detecting a signal of 13C-bicarbonate or the signal of 13C-bicarbonate and the signal of 13C-pyruvate where the said hyperpolarised 13C-pyruvate is selected from the group consisting of hyperpolarized 13C1-pyruvate, 13C1,2-pyruvate, 13C1,3-pyruvate or 13C1,2,3-pyruvate or any of their combination, the activity of PDH complex is determined.

EFFECT: use of the claimed method enables to determine reliably changes in activity of PDH complex in the subject to be examined.

11 cl, 7 dwg, 5 ex

FIELD: medicine.

SUBSTANCE: method for describing indications for choosing a conservative therapeutic approach to the patients suffered recurrent myocardial infarction involves pre-therapeutic patient's blood examination to analyse blood serum for oxidation-resistant lipoproteids (ORLs), to analyse erythrocytes for the pyruvic acid concentration (PAC), and if the ORL value is equal to 1.89 nmole MDA/mg of protein of β-lipoprotein and lower, while the PAC value is equal to 1.81 mmole/l and higher, the conservative therapy is added with the preparations of lipoic acid.

EFFECT: method is simple, has a broad information value and allows a more objective assessment of the patient's state and identifying a group of the patients in need of the conservative treatment and suffered recurrent myocardial infarction by the preparations of lipoic acid.

3 ex

FIELD: medicine.

SUBSTANCE: taken venous blood is separated into two samples. The first sample is stabilised with a solution of sodium citrate, the second one - with ethylene diamine tetraacetate. The first sample of whole blood is added with adenosine diphosphate as an aggregation inducer and tested for a peak amplitude of thrombocyte aggregation and a peak amplitude of adenosine triphosphate release profile by impedance method. The second sample is used to measure a fraction of thrombocytes and a fraction of blood corpuscles. It is followed by calculating a thrombocyte aggregation potential index by formula I=LmaxPTCΩmaxHTC100%; wherein Lmax is the peak amplitude of adenosine triphosphate release profile, Ωmax is the peak amplitude of thrombocyte aggregation, PCT is the fraction of thrombocytes, HTC is the fraction of blood corpuscles. If the value I is less than 0.5%, the low clinical effectiveness of the antiaggregant therapy is stated, and the value I being 1.5-2.5% shows the high effectiveness thereof.

EFFECT: improving the objective estimation of the clinical effectiveness of the antiaggregant therapy in the patients with acute coronary syndrome, and providing an opportunity for predicting the clinical course of the disease.

1 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine. A composition for stimulating the skin stem cell production containing interleukin-1 alpha and a dermatologically acceptable diluent or carrier.

EFFECT: invention provides improving the stem cell stimulation.

2 ex

FIELD: chemistry.

SUBSTANCE: method involves dissolving 855 mg of a crystalline hydrate of copper chloride (CuCl2·2H2O) in 100 ml of distilled water (concentration of Cu2+ ions in the prepared solution is 50 mmol/l) and adding 1 ml of the prepared solution to 100 ml of a standard reagent used in glucose oxidase test. The ascorbic acid oxidant used is copper chloride solution in end concentration in the glucose oxidase reagent of 500 mcmol/l.

EFFECT: method enables correct determination of glucose content.

1 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: workers' blood serum is analysed for interleukin 4, protein S-100β, protein S-100 autoantibodies, voltage-dependent Ca-channel autoantibodies, glutamate receptor autoantibodies, γ-aminobutyrate receptor autoantibodies, dopamine receptor autoantibodies; diagnostic coefficients F1 and F2 are calculated; if the value F1 is less than F2, the early changes of the nervous system are diagnosed for the chronic exposure to vinyl chloride; F1 more or equal to F2 enables stating the absence of any signs of the chronic exposure to vinyl chloride. The developed method may be used in the periodic medical screenings, medical examinations of workers to diagnose some occupational diseases.

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1 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention may be used to predict a developing myocardial dysfunction in the children with acute lymphoblastic leukemia (ALL) at different stages of polychemotherapy (PCT). The method involves the blood examination for the iron metabolism parameters, namely before the beginning of polychemotherapy (1) and after the induction of remission (2), blood serum ferritin, hepcidin and iron are evaluated in the patients; the derived values are inserted into the equations to calculate varying ECG, IMS, B(E-Ea) NT-pro-BNP after the completion of the intensive PCT course (3) and the total coefficient K is calculated by formula K=ECG3* IMS3* B(E-Ea)3* NT-pro-BNP3, wherein a probability of the myocardial dysfunction is stated by the total coefficient, namely: the coefficient K> 0.24 ensures predicting the developing cardiac complications, while K <0.24 show a lower risk of the cardiac complications.

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1 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method consists in determining a characteristic profile of a test sample of a human biological fluid. It is concentrated off-line. Biologically active substances are separated using complexing additives, and a 'reference' is determined. Steroid hormones are taken as the analysed biologically active substances. The hormones are separated by performed by reversed-phase HELC in gradient elution using a diode array detector. The steroid profiles are used to form a matrix of the analytical signal intensities and the retention factors of each steroid. Each sample is graphically imaged by method of principal components, and the graphical images are used to form 'reference' and deviation clusters. The 'reference' and deviation clusters are corrected by soft independent modelling of class analogy taken as a reference. The pathologies are diagnosed by an ability of the patient's image to come with a 'reference' or a deviation.

EFFECT: reliable diagnosis of the pathologies associated with adrenal cortical diseases.

6 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: what is presented is a method for prediction the efficacy of the anti-TNF therapy in the patients with rheumatoid arthritis on the basis of genetic typing the polymorphisms of TNF-alpha proinflammatory cytokine. The allelic polymorphism of the TNF-alpha gene promoter is studied in position 857. If the heterozygous state (genotype - 857ST) or the homozygous T allele carriers (genotype - 857TT) is identified, a high probability of the successful infliximab therapy is predicted. If identifying the homozygous allele C carrier in position - 857 of the TNF-alpha gene promoter (genotype - 857SS), a high probability of the failed infliximab therapy is predicted.

EFFECT: invention enables the rapid and effective prediction of the clinical outcome of the anti-TNF therapy in the patients with rheumatoid arthritis by one polymorph position.

2 tbl, 1 ex

FIELD: measurement equipment.

SUBSTANCE: penetrating radiation is generated, filtered and subsequently sent via a control object, and the passed-through radiation is registered, at the same time the generated penetrating radiation is gamma radiation of thulium Tm170 isotope, and for filtration tungsten or tantalum is used with efficient thickness from 0.2 to 0.5 mm.

EFFECT: reduced complexity of equipment and its energy intensity with simultaneous increase of sensitivity to defects.

3 dwg

FIELD: measurement equipment.

SUBSTANCE: sample is previously frozen, a frozen sample under conditions of negative temperature is put in contact with a frozen solution of a radio-opaque agent, upon completion of sample saturation, computer X-ray microtomography of the sample is carried out under negative temperatures, and by means of analysis of the produced computer tomographic image they detect spatial distribution and concentration of ice and/or gas hydrate inclusions of open and closed porosity, distribution of pores by size, specific surface in the sample.

EFFECT: higher accuracy of assessment of characteristics of non-consolidated porous media.

10 cl, 1 dwg

FIELD: physics.

SUBSTANCE: quartz monofractions are collected and annealed to temperature of 400°C, followed by excitation of X-ray luminescence, wherein X-ray luminescence is excited in the 370 nm band and the optical transmission coefficient is estimated using a graph of conformity between X-ray luminescence intensity in the 370 nm band and optical transmission coefficient values determined using a standard technique.

EFFECT: faster and more reliable preliminary evaluation of quality of quartz material.

1 tbl, 2 dwg

FIELD: chemistry.

SUBSTANCE: method involves preliminary concentration of trace elements from ultra-small samples of water and aqueous solutions. A microgranule of hydrophilic material is placed on a stretched radiotransparent hydrophobic film, a droplet of the analysed solution is collected and then deposited on said film to coat said microgranule. Said droplet is evaporated and the microgranule is then analysed by an X-ray fluorescent microanalyser with a focusing X-ray lens. Before placing the microgranule, the film is smeared with a liquid surfactant which is insoluble in water and aqueous solutions. 1-5% concentrated nitric acid or chloric acid is added to the droplet of the analysed solution or said droplet is taken from the analysed solution to which nitric acid or chloric acid has been added until achieving concentration thereof of 1-5%.

EFFECT: fewer analytical errors.

3 cl, 1 tbl, 8 dwg, 4 ex

FIELD: physics.

SUBSTANCE: double-spectrum illumination mode with separate selection of signals arising from radiation absorption in a background substance and signals arising from radiation absorption in overlapping layers of the background substance and inclusion substances is executed, wherein the X-ray exposure procedure is carried out not in one but two mutually perpendicular geometric projections, which enable mutual quantitative comparison of the mass thickness of the inclusion in one of the projections with the value of the linear dimension of that inclusion in the other projection and determine density of the inclusion substance from their ratio.

EFFECT: high probability of detecting hazardous inclusions and significant reduction of the probability of false alarm.

3 cl, 1 dwg

FIELD: chemistry.

SUBSTANCE: invention can be used in chemical industry. Lithium-iron phosphate having an olivine crystal structure has a composition expressed by the chemical formula (I) L1+aFe1-xMx(PO4-b)Xb (where M is selected from Al, Mg, Ti; X is selected from F, S, N; -0.5≤a≤+0.5; 0≤x≤0.5; and 0≤b≤0.1 ), contains 0.1-5 wt % Li3PO4 and does not contain or contains less than 0.25 wt % Li2CO3. Content of Li3PO4 in the lithium-iron phosphate increases electrochemical stability and ensures thermal safety and ion conductivity.

EFFECT: lithium-iron phosphate according to the present invention can be used as an active material for a positive electrode of a secondary lithium battery.

15 cl, 1 tbl, 5 dwg

FIELD: physics.

SUBSTANCE: cuvette for analysed samples has an outer casing which forms a sample storage reservoir; a directional filling valve lying in the top end of the outer casing and forming the top edge of the sample storage reservoir, a filling valve for receiving a sample during filling and preventing sample leakage during ventilation after filling; and a film which covers the bottom edge of the outer casing, also forming the bottom edge of the sample storage reservoir, and a film for holding samples in the focal region of the analysing device. At least one X-ray optical unit can be placed on the excitation and/or detection path, requiring matching with the focal region.

EFFECT: enabling formation of a cuvette with an accurately defined shape, which minimises contamination, reduces the probability of errors caused by operators, and which enables accurate positioning of the sample with respect to the X-ray analysis device.

20 cl, 9 dwg

FIELD: physics.

SUBSTANCE: apparatus for determining the fractional amount of each phase of a multiphase fluid medium includes an X-ray generator and a sample chamber configured to hold a sample of the fluid medium for analysis. The chamber is placed on the path of the output of radiation from the generator. A filter is placed on the radiation path between the output of the generator and the input of the sample chamber. A first radiation detector is placed on the radiation path from the sample chamber after passage of the radiation through the sample chamber. The thickness and material of the filter is selected to optimise resolution of radiation detected by the first detector based on variations in volume fractions of oil and water in the sample of the fluid medium when the gas fraction therein is about 90-100%.

EFFECT: high accuracy under conditions which include high fractional volume of gas in the analysed fluid medium.

13 cl, 6 dwg

FIELD: physics.

SUBSTANCE: fast neutron source is placed in a slowing unit; a γ-spectrometer employing a non-overload linear detection technique is used to detect instantaneous γ-quanta resulting from radiation capture of neutrons by nuclei of elements; calibration responses of separate elements making up the sample being identified are determined; apparatus spectra of γ-quanta are used to determine concentration of elements in the sample through weight coefficients of responses of elements, wherein chemical composition of the media is determined by: a priori determining the chemical composition - known chemical compounds - of the medium; determining calibration responses of compounds through the sum of calibration responses of elements making up the compounds; determining concentration of compounds and elements in the medium being identified; determining concentration of elements making up the compounds; establishing conformity of the obtained total concentration of elements of the medium, based on elements making up compounds of the medium being identified, concentration of elements of the medium obtained by identification decryption of only the elemental composition of the medium; in case of mismatch with a given error of the total concentration of elements of the medium, obtained by identification decryption based on elements making up the identified compounds of the medium, with concentration of elements of the medium, obtained by identification decryption of only the elemental composition of the medium, the procedure, starting from the a priori determining of the structure of compounds, is repeated.

EFFECT: faster elemental analysis, high resolution of identification of elements, high sensitivity of determining impurities in media.

3 cl, 13 dwg

Gamma-flaw detector // 2477463

FIELD: machine building.

SUBSTANCE: gamma-flaw detector includes a composite protection unit in the form of a stationary one-piece cylinder equipped with an eccentric cavity and a collimation outlet opening and corresponding to the eccentric cavity of a turning rotor equipped by means of a protective rod-shaped holder with an emission source in the eccentric channel with a hole coaxial to the collimation opening, and device for rotor turning through 180°. On the drive half-axis of the rotor there reinforced is a gear with external engagement, the end surface of which is formed with an arc-shaped groove concentrically to its constituent surface; the above groove provides the possibility of its discrete rotation together with the rotor in the azimuth angle range of 180°, which is strictly restricted with a limit stop corresponding to the groove profile and made in the form of a cylindrical pin statically reinforced in the housing. The gear is constantly adjacent kinematically and in a reduced manner (reduction of not less than 2) to a sector of a gear wheel, which is polarised by means of a spring and reinforced on the corresponding axis with possibility of pendulum movements transmitted to it with a rotor turning device through the gear and providing the polarised power fixation of the rotor in diametrically opposite discrete positions of emission and overlapping of a radiation beam.

EFFECT: increasing safety and improving reliability of protective properties considering sanitary norms of radiation security, improving accuracy and reliability of the control method.

4 cl, 4 dwg

FIELD: measurement equipment.

SUBSTANCE: method to detect solidity of liquid flow in a pipeline, when the liquid flow is exposed to electric field, the controlled flow is probed with an electromagnet wave, and the electromagnet wave passing through the flow is received. At the same time flow probing is carried out orthogonally to power lines of the electric field, amplitude of electric field of the elliptically polarised wave that passed through the liquid flow is measured, and the measured value of amplitude of the electric field of this wave determines solidity of the liquid flow in the pipeline.

EFFECT: simplified procedure of flow solidity measurement.

1 dwg

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