JP2006333783A - Biochemical reaction cartridge, biochemical rector, and biochemical reaction testing system using the biochemical reaction cartridge and the biochemical rector - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To solve the problems wherein a conventional biochemical cartridge has no method for measuring the amount of a specimen injected to a chamber for accumulating the specimen, and when the amount of the specimen in the cartridge is insufficient, the detection cannot be carried out in high reliability. <P>SOLUTION: The biochemical reaction cartridge has a plurality of chambers at least comprising the chamber for accumulating the specimen, a chamber for the chemical reaction, capable of accumulating a solution for carrying out a biochemical treatment, and a reaction chamber for carrying out the detection reaction of a target material in the biochemically treated specimen, and a plurality of passages communicating the chambers. The chamber for accumulating the specimen has the first detection means for detecting the amount of the specimen injected to the chamber for accumulating the specimen. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a biochemical reaction cartridge, a biochemical reaction device, and a biochemical reaction inspection system using the biochemical reaction cartridge and the biochemical reaction device.

  Many analyzers for analyzing a sample such as blood use an immunological method utilizing an antigen-antibody reaction or a method utilizing nucleic acid hybridization. For example, antibodies such as antibodies or antigens that specifically bind to the detected substance or antigens or single-stranded nucleic acids are used as probes, and immobilized on a solid surface such as fine particles, beads, glass plates, etc., and the detected substance and antigen-antibody reaction Alternatively, nucleic acid hybridization is performed. And using a labeling substance having a specific interaction carrying a labeling substance having high detection sensitivity such as an enzyme, a fluorescent substance, a luminescent substance, such as a labeled antibody, a labeled antigen or a labeled nucleic acid, An antigen-antibody compound or a double-stranded nucleic acid is detected to detect the presence or absence of a test substance or to quantify the test substance.

  As a development of these techniques, for example, US Pat. No. 5,445,934 discloses so-called DNA in which a large number of DNA (deoxyribonucleic acid) probes having different base sequences are arranged in an array on a substrate. An array is disclosed.

  Anal. Biochem. 270 (1), 103-111, 1999 discloses a method of preparing a protein array having a structure like a DNA array by arranging many kinds of proteins on a membrane filter. As described above, it has become possible to test a very large number of items at once by using a DNA array, a protein array, or the like.

  We also propose disposable biochemical reaction cartridges that carry out necessary reactions internally for the purpose of reducing sample contamination, improving reaction efficiency, miniaturizing equipment, and simplifying work in various sample analysis methods. Has been.

  For example, in Japanese Patent Application Laid-Open No. 11-509094, a plurality of chambers are arranged in a biochemical reaction cartridge including a DNA array, a solution is moved by differential pressure, and DNA in a sample is extracted or amplified inside the cartridge. Alternatively, a biochemical reaction cartridge that enables a reaction such as hybridization is disclosed.

  As a method for injecting a solution from the outside into such a biochemical reaction cartridge, there is a method using an external syringe pump or a vacuum pump. As a method for moving a solution inside a biochemical reaction cartridge, a method using gravity, capillary action, or electrophoresis is known. As a micro pump that is small and can be disposed inside the biochemical reaction cartridge, Japanese Patent No. 2832117 uses a heating element, Japanese Patent Application Laid-Open No. 2000-274375 uses a piezoelectric element, and special table. Japanese Patent Laid-Open No. 11-509094 discloses a diaphragm pump.

  From the viewpoint of preventing secondary infection and contamination, and from the viewpoint of ease of use, it is preferable to use a disposable cartridge that contains the necessary solution. However, there is a problem that the cartridge that contains the pump is expensive. Disposable biochemical reaction with a structure that allows a series of biochemical reactions to proceed without causing the solution to flow outside after the user injects the sample without moving the solution by the action of an external pump. Cartridge proposals have also been made.

  As a method for injecting a specimen such as blood into the biochemical reaction cartridge, for example, as disclosed in US Pat. No. 6,458,545, a method is disclosed in which an injection needle is attached to a blood collection tube containing a chip and blood is introduced into the collection part. ing.

Generally, a sample is collected from a subject using a syringe or a collection tube and a collection tube holder, and when transferred to a test tube, the examiner injects the sample using a syringe, its syringe, or pipette. .
US Pat. No. 5,445,934 Japanese National Patent Publication No. 11-509094 Japanese Patent No. 2832117 JP 2000-274375 A US Pat. No. 6,458,545 Lueking A, Horn M, Eickhoff H, Bussow K, Lehrach H, Walter G, Protein microarrays for gene expression and analysis screening, Anal. Biochem. 270 (1), 103-111, 1999

  However, the conventional biochemical cartridge has no method for measuring the amount of the sample injected into the chamber for storing the sample, and a method for detecting it even if the sample is mistakenly injected or less than the required amount. There was no.

  If the amount of the sample in the cartridge is insufficient, there is a problem that a highly reliable test cannot be performed. In particular, in a test using DNA from a sample, if the amount of the sample is insufficient, the necessary sample DNA cannot be sufficiently hybridized with a DNA probe, and a problem that a highly reliable test cannot be performed easily occurs.

  If the amount of specimen in the cartridge is too large, the specimen may overflow from the cartridge and cause contamination.

In order to achieve the above object, the present invention comprises at least a chamber for accumulating a specimen, a chemical reaction chamber capable of accumulating a solution for biochemical treatment, and a detection reaction of a target substance in the biochemically treated specimen. A biochemical reaction cartridge having a plurality of chambers composed of reaction chambers for performing and a flow path communicating between the plurality of chambers,
A biochemical reaction cartridge characterized in that the chamber for accumulating the sample includes a first detection means for detecting the amount of the sample injected into the chamber for accumulating the sample, A holding means for holding a biochemical reaction cartridge having a chamber for accumulating a specimen into which a specimen is injected at least from the outside; and a second detecting means for detecting the amount of the specimen injected into the biochemical reaction cartridge. A biochemical reaction device that performs a biochemical reaction using a biochemical reaction cartridge.

Furthermore, a chamber for accumulating at least an analyte, a chemical reaction chamber capable of accumulating a solution for biochemical treatment,
A plurality of chambers comprising a reaction chamber for performing a detection reaction of a target substance in a biochemically processed specimen;
A flow path communicating between a plurality of chambers;
A biochemical reaction cartridge comprising a first detection means for detecting the amount of the specimen injected into the chamber for storing the specimen;
Holding means for holding at least a biochemical reaction cartridge;
A biochemical reaction device having a second detection means for detecting the amount of the sample injected into the biochemical reaction cartridge,
A first detection means of a biochemical reaction cartridge mounted on a holding means of a biochemical reaction device for performing a biochemical reaction;
A biochemical reaction test system comprising: a determination unit that determines the amount of the specimen accumulated in the biochemical reaction cartridge using the second detection unit of the biochemical reaction device.

  As described above, according to the present invention, since the amount of the sample injected into the chamber for storing the sample can be detected, the biochemical reaction can be started in a state where a desired amount of the sample exists. Therefore, the reliability of the inspection can be ensured.

  The present invention detects the amount of a sample injected into a chamber for storing a sample in a biochemical cartridge.

The amount of sample injected into the chamber that stores the biochemical reaction cartridge sample is:
1. 1. A transparent window is formed in which the specimen accumulated in the specimen accumulation chamber can be monitored from the side of the biochemical reaction cartridge. 2. An exposed electrode is provided in the chamber for accumulating the specimen. It can be monitored by measuring the weight of the biochemical reaction cartridge.

  A biochemical reaction cartridge used in the present invention and a biochemical reaction apparatus that performs biochemical reaction and inspection using the biochemical reaction cartridge will be described in detail with reference to the drawings.

  1 and 2 are biochemical reaction cartridges used in this embodiment, and FIG. 3 is a biochemical reaction apparatus using the cartridge shown in FIGS.

  FIG. 1 is an external view of the biochemical reaction cartridge 1. At the top of the biochemical reaction cartridge 1, a sample inlet 2 for injecting a sample such as blood using a syringe or the like is provided on the side surface of the biochemical reaction cartridge 1 (the opposite surface has the same configuration). In order to move the solution inside, a plurality of nozzle inlets 3 are provided to insert a nozzle and pressurize or depressurize it. A rubber cap is fixed to the sample inlet 2 and each nozzle inlet 3.

  As a material constituting the main body of the biochemical reaction cartridge 1, polymethyl methacrylate (PMMA), acrylonitrile-butadiene-styrene (ABS) copolymer, polystyrene, polycarbonate, polyester, polyvinyl chloride, or the like is preferably used. .

  FIG. 2 shows a plan cross-sectional view of the biochemical reaction cartridge 1, wherein ten nozzle inlets 3a to 3j are provided on one side surface, and ten nozzle inlets 3k to 3t are provided on the opposite side surface. Yes. Each nozzle inlet 3a-3t is connected to the chamber 5 which is a place which stores a solution or a place which reacts via the air flow paths 4a-4t through which each air flows.

  However, since the nozzle inlets 3n, 3p, 3q, and 3s are not used in the process of the present embodiment, they are not communicated with the chamber 5 and are reserved. That is, the nozzle inlets 3a to 3j are communicated with the chambers 5a to 5j via the flow paths 4a to 4j, and the opposite nozzle inlets 3k, 3l, 3m, 3o, 3r, and 3t are respectively connected to the flow paths 4k and 4l. 4m, 4o, 4r, and 4t, and communicated with chambers 5k, 5l, 5m, 5o, 5r, and 5t.

  The specimen inlet 2 communicates with the chamber 7, the chambers 5 a, 5 b, 5 c and 5 k communicate with the chamber 7, the chambers 5 g and 5 o communicate with the chamber 8, and the chambers 5 h, 5 i, 5 j, 5 r and 5 t communicate with the chamber 9. . Further, the chamber 7 communicates with the chamber 8 via the flow path 10, and the chamber 8 communicates with the chamber 9 via the flow path 11. Chambers 5d, 5e, 5f, 5l, and 5m are communicated with the flow path 10 through flow paths 6d, 6e, 6f, 6l, and 6m, respectively.

  In addition, a square hole is formed in the bottom surface of the chamber 9, and several tens to hundreds of thousands of different types of DNA probes are arranged on the solid phase surface such as a glass plate having a size of about square inch. A DNA microarray 12 is attached with the probe surface facing up.

  By performing a hybridization reaction with the sample DNA using this DNA microarray 12, a large number of genes can be examined at once. In addition, these DNA probes are regularly arranged in a matrix, and the DNA probe information can be easily extracted from the position information (number of rows / columns) of the DNA probes on the DNA microarray 12. In addition to infectious disease viruses, bacteria, and disease-related genes, genes to be tested include individual polymorphisms.

  FIG. 3 shows a schematic diagram of a biochemical reaction apparatus 30 that controls the movement of the solution in the biochemical reaction cartridge 1 and various reactions. The biochemical reaction cartridge 1 is set on the table 13. On the table 13, an electromagnet 14 that is activated when DNA or the like from the specimen is extracted in the biochemical reaction cartridge 1, and temperature control is performed when the DNA from the specimen is amplified by a method such as PCR (Polymerase Chain Reaction). Temperature control when performing hybridization between the amplified sample DNA and the DNA probe on the DNA microarray in the biochemical reaction cartridge 1 and washing the sample DNA that has not been hybridized Peltier elements 16 are arranged to connect to a control unit 17 that controls the entire processing apparatus.

  On both sides of the table 13, there are electric syringe pumps 18, 19 and pump blocks in which ten pump nozzles 20, 21 are provided on each side, with an inlet / outlet for discharging or sucking air by these pumps 18, 19. 22 and 23 are arranged. A plurality of electric switching valves (not shown) are arranged between the electric syringe pumps 18 and 19 and the pump nozzles 20 and 21, and are connected to the controller 17 together with the pumps 18 and 19.

  Moreover, the control part 17 is connected to the input part 24 which an inspector inputs. The control unit 17 performs control to selectively open the pump nozzles 20 and 21 one by one with respect to the electric syringe pumps 18 and 19 or to close all the pump nozzles among the ten nozzles on one side. It has become.

  In FIG. 3, an illumination light source 25 and a detection unit 27 for detecting the amount of the sample contained in the biochemical reaction cartridge 1 are provided in the biochemical reaction device 30, and the illumination light source 25 and the detection unit 27 are included in the control unit 17. However, the illumination light source 25 may be provided if the detection unit is not an optical detection unit, and will be described later when a two-dimensional sensor is used even if the detection unit is an optical detection unit. There is a case where it is not necessary to provide it.

  In the chambers 5a and 5b, a first hemolytic agent containing EDTA that destroys the cell wall and a second hemolytic agent containing a protein denaturant such as a surfactant are accumulated. Silica-coated magnetic particles to which DNA is adsorbed are accumulated in the chamber 5c, and the first and second extraction detergents used for purifying the DNA at the time of DNA extraction in the chambers 5l and 5m. Is accumulated.

  The chamber 5d contains an eluate composed of a low-concentration salt buffer that elutes DNA from magnetic particles, and the chamber 5g contains primers necessary for PCR, polymerase, dNTP solution, buffer, Cy-3dUTP containing a fluorescent agent, and the like. The mixture is filled. The chambers 5h and 5j have a cleaning agent containing a detergent for washing the fluorescently labeled sample DNA and fluorescent label, and the chamber 5i has the inside of the chamber 9 including the DNA microarray 12 dried. Alcohol for accumulating is accumulated.

  The chamber 5e is a chamber for collecting dust other than blood DNA, the chamber 5f is a chamber for collecting waste liquids of the first and second extraction detergents in the chambers 5l and 5m, and the chamber 5r is for the first and second detergents. Chambers 5k, 5o, and 5t are blank chambers provided to prevent the solution from flowing into the nozzle inlet.

(First embodiment)
FIG. 8 is a side view of the biochemical reaction cartridge used in the first embodiment as seen from the direction of arrow A shown in FIG. At least a part of the side surface of the arrow A in the chamber 7 in which the specimen is accumulated becomes a window made of an optically transparent material X so that the quantity of the specimen can be optically detected from the outside of the biochemical cartridge. Therefore, the amount of the specimen in the chamber 7 can be confirmed. The transparent material X is formed to constitute the chamber 7 and the side wall of the biochemical reaction cartridge 1.

  In the transparent member X, the biochemical reaction cartridge 1 is positioned at a position that becomes a horizontal plane when a desired amount of the sample to be injected into the chamber 7 is placed in a state where the biochemical reaction cartridge 1 is placed substantially horizontally so that the sample inlet 2 is on the upper surface. A reference line T is provided.

  For example, when blood is used as a specimen, when the tester injects blood with the syringe penetrating the rubber cap of the specimen inlet 2 with the biochemical reaction cartridge 1 placed so that the specimen inlet 2 is on the upper surface, Blood is injected into the chamber 7.

  FIG. 8 shows an example in which the position reference line T is formed at one place. When the position reference line T is formed at one place, a position corresponding to a horizontal plane when a substantially desired amount of specimen is injected into the chamber 7 or a necessary minimum quantity (lower limit quantity) of specimen is placed in the chamber 7. It is preferable to form the position reference line T at a position or the like corresponding to the horizontal plane when injected.

  In FIG. 9, the reference line T and the maximum amount (upper limit amount) of the sample are injected into the position indicating the horizontal plane when the necessary minimum amount (lower limit amount) of the sample is injected as the position reference line (reference line). This is an example in which two position reference lines of a reference line T ′ are provided at a position indicating a horizontal plane.

  The position reference line can be provided in addition to the positions described with reference to FIGS.

  A window composed of a transparent material X of the biochemical reaction cartridge 1 from the illumination light source 25 of the biochemical reaction device in order to detect that a desired amount of the sample is injected into the chamber 7 in the chamber 7 for accumulating the sample. Irradiate the light. Next, the window formed of the transparent material X of the biochemical reaction cartridge 1 is monitored by the detection unit 27 formed of a light receiving element. As the light receiving element constituting the detection unit 27, a one-dimensional photosensor such as a light receiving diode or a two-dimensional photosensor such as a CCD (Charge Coupled Devices) or a CMOS sensor can be used.

  When detecting the amount of the specimen based on the position reference line, it is preferable to use a two-dimensional optical sensor such as a CCD or a CMOS sensor as the detection unit 27. A position reference line can be used as a reference from an image monitored by a two-dimensional optical sensor such as a CCD or CMOS sensor. In this case, as shown in FIG. 9, when two types of reference lines indicating the maximum amount and the minimum amount of the sample are provided as the position reference lines, the sample is between the two types of position reference lines. In this case, since it can be determined that a desired amount of specimen is accumulated in the chamber 7, it can be easily determined.

  When the position reference line is provided at one position at a position corresponding to the horizontal plane when a substantially desired amount of the sample is injected into the chamber 7, the distance between the position reference line and the sample surface is Whether or not it is determined that a desired amount of specimen is accumulated in the chamber 7 can be stored in advance.

  Further, even if the position reference line is not provided, the determination can be made by storing the positions of the minimum amount and the maximum amount of the sample in advance.

  When a one-dimensional optical sensor such as a light receiving diode is used as the detection unit 27, it is not necessary to provide the position reference line shown in FIGS. 8 and 9 in the window, and detection is performed using a change in the reflectance of the window light. be able to. In this case, in order to determine that a desired amount of sample has been injected into the chamber 7, the control unit 17 has a reflectance 1 of the window when the minimum necessary sample is injected into the chamber 7 and the chamber 7. The reflectance 2 of the window when the maximum specimen is injected is stored, and the reflectance of the measured window is compared with reflectance 1 and reflectance 2, and a desired amount of specimen is injected into the chamber 7. Can be determined.

  When the reflectance is lowered by injecting the specimen, the relationship of reflectance 1 <measured reflectance <reflectance 2 is satisfied. When the reflectance is raised by injecting the specimen, reflectance 1> measurement. When the relationship of reflectivity> reflectivity 2 is satisfied, it may be determined that a desired amount of specimen is accumulated in the chamber 7. When measuring reflectance, it is preferable to provide the illumination light source 25.

  When a CCD or CMOS sensor with high photosensitivity is used, the illumination light source 25 may not be provided.

  When the biochemical reaction cartridge 1 in which the specimen is injected into the chamber 7 is placed on the table 13 of the biochemical reaction apparatus 30 and the processing start command is issued from the input unit 24, the specimen amount check is started. First, the illumination light source 25 illuminates the portion formed of the transparent material X on the side surface of the cartridge, the detection unit 26 captures the image of the side X portion, and the acquired image is sent to the control unit 17.

  At this time, since the side X portion is transparent, the amount of the specimen B can be confirmed. Then, it is preferable that the control unit 17 has a determination unit that compares the amount of the specimen B with a preset reference amount T from the acquired image and determines whether or not to perform a biochemical reaction process.

  When the sample amount is larger or smaller than the desired amount, it is preferable that the determination unit transmits a signal to a warning generation unit (not shown) and generates a warning from the warning unit.

  As a warning means, a sound can be generated or a warning can be generated by light. Further, when the sample amount is larger or smaller than the desired amount, the determination unit sends a signal for stopping the next operation to the control unit 17, and the control unit 17 can stop the operation based on this signal. .

  It is preferable to perform the warning and the work stop simultaneously.

  In the present embodiment, an example in which the illumination light source 25 and the detection unit 26 are provided in the biochemical reaction device 30 has been described. However, the biochemical reaction cartridge 1 when the specimen is injected is placed on a table (not shown). An illumination light source (not shown) and a detection unit (not shown) can also be provided. In this case, it is preferable that a means for receiving the signal from the detection unit and determining the amount of the sample is connected to the detection unit.

  It is preferable to provide a determination unit that determines the amount of the sample based on the result of the detection unit and a warning unit that issues a warning based on the result of the determination unit.

(Second Embodiment)
Also, as shown in FIG. 10, if a weight sensor 27 is provided on the table 13 and the output of the weight sensor 27 is connected to the control unit 17 by means not shown, the sample-injected cartridge is placed on the weight sensor 27. If set to, the sample amount can be detected by weight.

  In the present embodiment also, it is preferable to have a determination means for determining whether or not to perform a biochemical reaction process.

  When the sample amount is larger or smaller than the desired amount, it is preferable that the determination unit transmits a signal to a warning generation unit (not shown) and generates a warning from the warning unit.

  As a warning means, a sound can be generated or a warning can be generated by light. Further, when the sample amount is larger or smaller than the desired amount, the determination unit sends a signal for stopping the next operation to the control unit 17, and the control unit 17 can stop the operation based on this signal. .

  It is preferable to perform the warning and the work stop simultaneously.

  As the weight sensor, an electrostatic gravity sensor, a weight sensor using a strain sensor, or the like can be used. When a gravity sensor is used, a conventional cartridge can be used as the biochemical reaction cartridge, so that it is not necessary to newly manufacture the biochemical reaction cartridge.

  Also in the present embodiment, as in the first embodiment, a gravity sensor serving as a detection means and a determination means are provided on a table on which the biochemical reaction cartridge 1 for injecting a specimen is placed, and the gravity sensor The amount of the sample can be determined by receiving the signal.

  Warning means for issuing a warning in response to the result of the determination means can be provided.

(Third embodiment)
Alternatively, as another method, as shown in FIG. 11, for example, an electrode 28 can be provided in the chamber 7. The electrode 28 that is paired with the electrode 28 provided at a position on the side surface of the chamber 7 where the desired amount of sample is provided is the same height as the electrode 28 formed on the surface facing the sample inlet 2 of the chamber 7 or on the side surface of the chamber 7. It is preferable to provide it at a lower position. When the electrode 28 is disposed at a position indicating the minimum amount (lower limit amount) necessary for the sample and a position indicating the maximum amount (upper limit amount) of the sample, the electrode 28 paired with these electrodes 28 is provided in the chamber 7. The electrode 28 is preferably formed at the same height as or lower than the electrode 28 formed at a lower position of the electrode 28 formed on the surface or side surface facing the specimen inlet 2.

  Although an example in which the electrode 28 is formed at one or two positions on the side surface is shown, the electrode on the side wall is not limited to the present embodiment.

  One end of the electrode 28 formed so as to be exposed to the chamber 7 is formed so as to be exposed on the outer surface of the biochemical reaction cartridge (not shown). These terminals are connected to the detection unit 27 provided in the biochemical reaction device 30. In this connection, the amount of the specimen can be detected by applying a voltage between the pair of electrodes 28 and the electrode formed on the side wall of the chamber 7 and detecting the current flowing between the electrodes.

  In this embodiment also, it is preferable to have a determination means for determining whether or not to perform biochemical reaction processing.

  When the sample amount is larger or smaller than the desired amount, it is preferable that the determination unit transmits a signal to a warning generation unit (not shown) and generates a warning from the warning unit.

  As a warning means, a sound can be generated or a warning can be generated by light. Further, when the sample amount is larger or smaller than the desired amount, the determination unit sends a signal for stopping the next operation to the control unit 17, and the control unit 17 can stop the operation based on this signal. .

  It is preferable to perform the warning and the work stop simultaneously.

  Also in the present embodiment, as described in the first embodiment and the second embodiment, the warning is based on the result of the determination unit and the determination unit that determine the amount of the sample based on the signal of the detection unit. Needless to say, it is possible to provide a warning means that emits the above. Further, these detection means, determination means, and warning means can be provided on a table on which the biochemical reaction cartridge 1 is placed when the specimen is injected. .

  As described in the above embodiment, in the present invention, the amount of the sample injected into the chamber 7 for storing the sample can be optimized, so that the number of amplifications for securing the necessary amount of DNA is constant. Even so, it is possible to perform highly reliable measurement. Furthermore, it is not necessary to collect more specimen than necessary from the examiner, and the burden on the subject can be reduced.

  From the viewpoint of preventing the possibility that the specimen overflows from the chamber 7 for accumulating the specimen, it can be said that it is sufficiently worth detecting not only the minimum required quantity but also the upper limit of the specimen quantity.

(Fourth embodiment)
FIGS. 4A and 4B are flowcharts for explaining the sample amount detection step.

  First, in FIG. 4A, in step S1, an image of the chamber portion into which the sample has been injected is captured by the detection unit 26 to detect the sample amount, and in step S2, it is determined whether the sample amount satisfies a predetermined amount. In the case of OK, the reaction process is started in step S3. In the case of NG, a warning is issued in step S4, and the process is stopped.

  On the other hand, FIG. 4 (2) shows a case where the specimen is not injected by the examiner, but if the examiner sets the specimen in the biochemical reaction apparatus, the specimen is injected into the cartridge by the biochemical reaction apparatus. ing. First, sample injection is performed in step S1, the sample amount is detected in step S2, and it is determined in step S3 whether the sample amount satisfies a predetermined amount. In the case of OK, the reaction process is started in step S4. In the case of NG, the process returns to step S1 to reinject the specimen.

  After the above sample amount check process is completed, the process proceeds to the reaction processing process.

  Next, when the biological reaction pump blocks 22 and 23 are moved in the direction of the arrow in FIG. 3, the pump nozzles 20 and 21 are inserted into the nozzle inlets 3 on both sides of the biochemical reaction cartridge 1 through the rubber caps.

  Thereafter, a biochemical reaction process is performed according to the processing procedure in the processing apparatus as shown in FIG. First, in step S1, the control unit 17 opens only the nozzle inlets 3a and 3k, discharges air from the electric syringe pump 18, sucks air from the pump 19, and puts the first hemolytic agent in the chamber 5a into the blood. Pour into the chamber 7. At this time, depending on the viscosity of the hemolytic agent and the resistance of the flow path, if the suction of the air from the pump 19 is controlled to start 10 to 200 milliseconds after the start of the discharge of the air from the pump 18, The solution does not jump out at the beginning of the flowing solution, and the solution flows smoothly.

  In this way, by controlling the pressurization and decompression by shifting the timing of air supply and suction, the solution can flow smoothly, but the suction of air by the electric syringe pump 19 If fine control such as linear increase from the start is performed, the solution can flow more smoothly. The same applies to the movement of the following solutions.

  The control of the air supply can be easily realized by using the electric syringe pumps 18 and 19. Only the nozzle inlets 3 a and 3 o are opened, and the discharge and suction of the air are alternately repeated by the pumps 18 and 19. The operation of flowing the solution through the flow path 10 and then returning to the flow path 10 is repeated to perform stirring. Alternatively, the air may be continuously discharged from the pump 19 and stirred while generating bubbles.

  FIG. 6 is a cross-sectional view through the chambers 5a, 7, and 5k shown in FIG. 2, and the pump nozzle 20 is inserted into the nozzle inlet 3a for pressurization, and the pump nozzle 21 is inserted into the nozzle inlet 3k for depressurization. This shows that the first hemolytic agent flows into the chamber 7 containing blood.

  In FIG. 5, next, in step S2, only the nozzle inlets 3b and 3k are opened, and the second hemolytic agent in the chamber 5b is poured into the chamber 7 in the same manner. In step S3, only the nozzle inlets 3c and 3k are opened, and the same Then, the magnetic particles in the chamber 5 c are poured into the chamber 7. In steps S2 and S3, stirring is performed in the same manner as in step S1. In step S3, the DNA obtained by dissolving the cells in steps S1 and S2 adheres to the magnetic particles.

  In step S4, the electromagnet 14 is turned on, only the nozzle inlets 3e and 3k are opened, the air is discharged from the electric syringe pump 19, the air is sucked from the pump 18, and the solution in the chamber 7 is moved to the chamber 5e. During this movement, the magnetic particles and DNA are captured on the electromagnet 14 in the flow path 10. The suction and discharge of the pumps 18 and 19 are alternately repeated, and the solution is reciprocated twice between the chambers 7 and 5e, thereby improving the DNA capture efficiency. Furthermore, if the number of times is increased, the capture efficiency can be further increased, but extra processing time is also required.

  In this way, DNA is captured in a flowing state on a small channel having a width of about 1 to 2 mm and a height of about 0.2 to 1 mm using magnetic particles, so that it can be captured extremely efficiently. Can do. The same applies when the capture target substance is RNA or protein.

  Next, in step S5, the electromagnet 14 is turned off, only the nozzle inlets 3f and 3l are opened, the air is discharged from the electric syringe pump 19, the air is sucked from the pump 18, and the first extraction cleaning liquid in the chamber 5l is removed. Move to chamber 5f. At this time, the magnetic particles and DNA captured in step S4 are moved together with the extraction washing liquid to perform washing. After reciprocating twice in the same manner as in step S4, the electromagnet 14 is turned on, and reciprocating twice in the same manner to collect the magnetic particles and DNA on the electromagnet 14 in the flow path 10 and return the solution to the chamber 5l. .

  In step S6, using the nozzle inlets 3f and 3m, the second extraction cleaning liquid in the chamber 5m is further cleaned by performing the same process as in step S5. In step S7, with the electromagnet 14 turned on, only the nozzle inlets 3d and 3o are opened, air is discharged from the electric syringe pump 18, and air is sucked from the pump 19, whereby the eluate from the chamber 5d is supplied to the chamber 8. Moving.

  At this time, the magnetic particles and DNA are separated by the action of the eluate, and only the DNA moves to the chamber 8 together with the eluate, and the magnetic particles remain in the flow path 10. In this way, DNA extraction and purification are performed. Since the chamber containing the extracted washing liquid and the chamber containing the waste liquid after washing are prepared, DNA can be extracted and purified in the biochemical reaction cartridge 1.

  Next, in step S8, only the nozzle inlets 3g and 3o are opened, air is discharged from the electric syringe pump 18, the air is sucked from the pump 19, and the PCR drug in the chamber 5g is poured into the chamber 8. Further, only the nozzle inlets 3g and 3t are opened, and the pumps 18 and 19 alternately repeat the discharge and suction of air to flow the solution in the chamber 8 through the flow path 11, and then repeat the operation of returning to the subsequent stirring. Then, after controlling the Peltier element 15 to keep the solution in the chamber 8 at a temperature of 96 ° C. for 10 minutes, the process of 96 ° C. · 10 seconds, 55 ° C. · 10 seconds, 72 ° C. · 1 minute is repeated 30 times. The amplified DNA is amplified by PCR.

  In step S9, only the nozzle inlets 3g and 3t are opened, air is discharged from the electric syringe pump 18, the air is sucked from the pump 19, and the solution in the chamber 8 is moved to the chamber 9. Further, the Peltier element 16 is controlled to perform hybridization by holding the solution in the chamber 9 at 45 ° C. for 2 hours. At this time, the pumps 18 and 19 alternately repeat the discharge and suction of air, move the solution in the chamber 9 to the flow path 6t, and repeat the operation of returning to the subsequent, and the hybridization proceeds.

  Next, in step S10, while maintaining the same 45 ° C., this time, only the nozzle inlets 3h and 3r are opened, air is discharged from the electric syringe pump 18, air is sucked from the pump 19, and the solution in the chamber 9 is discharged. The first cleaning liquid in the chamber 5 h is poured into the chamber 5 r through the chamber 9 while moving to the chamber 5 r. The suction and discharge of the pumps 18 and 19 are alternately repeated to reciprocate the solution twice between the chambers 5h, 9, and 5r, and finally return to the chamber 5h. In this way, the fluorescently labeled sample DNA and the fluorescent label that have not been hybridized are washed.

  FIG. 7 is a cross-sectional view through the chambers 5h, 9, and 5r of FIG. 2. The pump nozzle 20 is inserted into the nozzle inlet 3h and pressurized, the pump nozzle 21 is inserted into the nozzle inlet 3r, and the pressure is reduced. The first cleaning liquid flows through the chamber 9 into the chamber 5r.

  In FIG. 5, in step S11, while maintaining the same 45 ° C., the second cleaning liquid in the chamber 5j is further cleaned through the same process as in step S10 using the nozzle inlets 3j and 3r. return. Thus, since the chambers 5h and 5j containing the cleaning liquid and the chamber 5r containing the waste liquid after the cleaning are prepared, the DNA microarray 12 can be cleaned in the biochemical reaction cartridge 1.

  In step S12, only the nozzle inlets 3i and 3r are opened, air is discharged from the electric syringe pump 18, the air is sucked from the pump 19, and the alcohol in the chamber 5i is moved through the chamber 9 to the chamber 5r. Thereafter, only the nozzle inlets 3i and 3t are opened, air is discharged from the pump 18, and air is sucked from the pump 19 to dry the inside of the chamber 9.

  When the inspector operates a lever (not shown), the pump blocks 22 and 23 move away from the biochemical reaction cartridge 1, and the pump nozzles 20 and 21 are detached from the nozzle inlet 3 of the biochemical reaction cartridge 1. Then, the examiner inserts the biochemical reaction cartridge 1 into a well-known scanner such as a scanner for DNA microarray to perform measurement and analysis.

  As described above, in the present embodiment, the sample amount is checked from the side of the cartridge. However, the entire cartridge or the vicinity of the chamber 7 constitutes a transparent member, and the extent of the sample is observed from the top or bottom surface of the cartridge. It is also possible to check the sample amount.

It is a perspective view of the biochemical reaction cartridge of an embodiment. It is a plane sectional view of a biochemical reaction cartridge. It is a block block diagram of the processing apparatus which controls the movement of a solution in a biochemical reaction cartridge, and various reaction. It is a flowchart figure of a sample detection processing procedure. It is a flowchart figure of a biochemical reaction processing procedure. It is a longitudinal cross-sectional view of some chambers. It is a longitudinal cross-sectional view of another one part chamber. It is the figure which showed the minimum of the sample required amount. It is the figure which showed the upper and lower limits of the required amount of the specimen. It is a processing apparatus figure at the time of using a weight detection sensor. It is a figure of the biochemical reaction cartridge which has arrange | positioned an electrode.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Biochemical reaction cartridge 2 Specimen inlet 3 Nozzle inlet 4 Air flow path 5, 7-9 Chamber 6, 10, 11 Flow path 12 DNA microarray 13 Table 17 Control part 18, 19 Electric syringe pump 20, 21 Pump nozzle 22, 23 Pump block 24 Input unit 25 Illumination 26 Detection unit 27 Weight sensor 28 Electrode 30 Biochemical reaction device X Transparent material B Sample T Reference line T 'Reference line

Claims (14)

A plurality of chambers including at least a specimen accumulation chamber, a chemical reaction chamber capable of accumulating a solution for biochemical treatment, and a reaction chamber for detecting a target substance in the biochemically treated specimen And a biochemical reaction cartridge having a flow path communicating between the plurality of chambers,
A biochemical reaction cartridge characterized in that the chamber for accumulating the specimen comprises first detection means for detecting the amount of the specimen injected into the chamber for accumulating the specimen.   The first detection means is a transparent region formed on a side surface of the biochemical reaction cartridge and made of a transparent member which is at least a part of a side wall of a chamber for accumulating the specimen. The biochemical reaction cartridge as described.   The biochemical reaction cartridge according to claim 2, wherein a reference line is formed in the transparent region.   The first detection means is at least two or more electrodes with one end exposed on the sample accumulation side of the chamber for accumulating the sample and the other end exposed on the outer surface of the biochemical reaction cartridge. The biochemical reaction cartridge according to claim 1.   At least a holding unit that holds a biochemical reaction cartridge having a chamber for storing a sample into which a sample is injected from the outside, and a second detection unit that detects the amount of the sample injected into the biochemical reaction cartridge. A biochemical reaction apparatus for performing a biochemical reaction using the biochemical reaction cartridge. Determination means for determining the amount of the sample based on the information of the second detection means;
6. The biochemical reaction device according to claim 5, further comprising warning means for issuing a warning based on the determination by the determination means. Determination means for determining the amount of the sample based on the information of the second detection means;
6. The biochemical reaction apparatus according to claim 5, further comprising a control unit that prohibits a subsequent operation based on the determination by the determination unit. Determination means for determining the amount of the sample based on the information of the second detection means;
Warning means for issuing a warning based on the determination by the determination means;
6. The biochemical reaction apparatus according to claim 5, further comprising a control unit that prohibits a subsequent operation based on the determination by the determination unit.   The biochemical reaction apparatus according to claim 5, wherein the second detection means is an optical means.   6. The chemical reaction apparatus according to claim 5, wherein the second detection means is an electrical means.   The biochemical reaction apparatus according to claim 5, wherein the second detection means is a weight detection means. A chamber for accumulating at least an analyte, a chamber for chemical reaction capable of accumulating a solution for biochemical treatment,
A plurality of chambers comprising a reaction chamber for performing a detection reaction of a target substance in a biochemically processed specimen;
A flow path communicating between the plurality of chambers;
A biochemical reaction cartridge comprising a first detection means for detecting the amount of the specimen injected into the chamber for accumulating the specimen;
Holding means for holding at least the biochemical reaction cartridge;
A biochemical reaction device having a second detection means for detecting the amount of the specimen injected into the biochemical reaction cartridge,
The first detection means of the biochemical reaction cartridge mounted on the holding means of a biochemical reaction apparatus that performs the biochemical reaction;
A biochemical reaction inspection system comprising: a determination unit that determines the amount of the specimen accumulated in the biochemical reaction cartridge using the second detection unit of the biochemical reaction device.   The first detection means is a transparent region formed on a side surface of the biochemical reaction cartridge and made of a transparent member which is at least a part of a side wall of the chamber for accumulating the specimen, and the second detection means is optical. The biochemical reaction inspection system according to claim 12, wherein the biochemical reaction inspection system is a specific means. The first detection means is at least two or more electrodes, one end of which is exposed on the side where the specimen is stored in the chamber for storing the specimen, and the other end is exposed on the outer surface of the biochemical reaction cartridge; The biochemical reaction inspection system according to claim 12, wherein the second detection means detects an electrical characteristic of the other end of the two or more electrodes.

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