JP2006122041A - Chemical analyzer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a chemical analyzer inhibiting hindering reaction characteristics such as a reaction liquid temperature fall just after enzyme addition, thus enabling stable detection. <P>SOLUTION: This chemical analyzer is accommodated with a structure that is introduced with a sample and holds a reagent to be reacted with the sample and is equipped with a mechanism to detect the sample after reacted with the reagent. In this analyzer, after extracting biomaterial(s) from the sample in the structure or during and after the extraction, a fluid in the ambient space of the structure is controlled at the optimum temperature for an enzyme. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a chemical analyzer that extracts and detects a specific chemical substance in a liquid sample.

  As a chemical analysis apparatus for extracting and analyzing a specific chemical substance such as a nucleic acid from a sample containing a plurality of chemical substances, Japanese Patent Publication No. 2001-527220 discloses an integrated fluid operation cartridge. In this apparatus, a reagent such as a lysing solution, a washing solution, or an eluent, and a capture component for capturing nucleic acid are provided inside the integrated cartridge. After injecting a sample containing nucleic acid into the cartridge, the sample and the eluent are mixed. And pass through the capture component, further pass the cleaning liquid through the capture component, further pass the eluent through the capture component, and contact the eluent after passing through the capture component with the PCR reagent to the reaction chamber. It is flowing. And the content of the heating using a thin film heater is disclosed as a temperature control means.

  In Japanese Patent Publication No. 2003-502656, an apparatus using a rotating disk and quantifying a sample using centripetal force and using a nucleic acid PCR amplification method is proposed. A structure using temperature control means for setting the denaturation temperature, annealing temperature, and elongation temperature in the PCR amplification method is disclosed in the rotating disk.

Special table 2001-527220 gazette Special table 2003-502656 gazette

  Both of the conventional techniques described in the above Japanese translations of PCT publication Nos. 2003-502656 and 2003-502656 use a nucleic acid amplification method in a PCR amplification method that repeats a temperature cycle. In the PCR amplification method, the nucleic acid is amplified according to the number of times by repeating the temperature cycle, for example, 95, 55, and 72 ° C. The prior literature merely discloses temperature control of the reaction liquid to a desired temperature based on this. No consideration is given to temperature control for improving the reaction characteristics or the temperature control system configuration. Also, temperature control for improving reaction characteristics during amplification is not considered.

  Accordingly, an object of the present invention is to provide a chemical analyzer that solves at least one of the above-described problems.

In order to solve the above problems, the present invention has the following aspects.
(1) A chemical analyzer including a structure in which a sample containing a biological material is introduced, a structure that holds a reagent to be reacted with the sample is accommodated, and includes a detection mechanism for the sample after reacting with the reagent, The fluid surrounding the structure is controlled to an appropriate reaction temperature of the reaction reagent after extraction of the biological material in the sample from the structure or during the extraction. Specifically, at least one of the reagents is a reagent containing an enzyme, a mechanism for extracting a biological material from the sample, a mechanism for supplying the reagent containing the enzyme to the extracted biological material, and the structure A temperature control mechanism that controls the temperature of the body, and the enzyme is contained by the temperature control mechanism after the step of extracting the biological material is started and before the reagent containing the enzyme is reacted. The analyzer is controlled to raise the temperature of the reagent.

The biological material is, for example, a nucleic acid. In addition, what is contained in biological samples, such as DNA, RNA, or protein, can be considered.
(2) In the chemical analyzer of (1), the biological material supplied with the enzyme is kept at a predetermined temperature for a predetermined time, and the biological material after the heat is controlled to be detected by the detection mechanism, In the step of raising the temperature, the chemical analyzer is controlled so that the reagent containing the enzyme approaches the heat retention temperature.

  For example, the predetermined temperature is a heat retention temperature such as an optimum temperature. Generally, the temperature is higher than room temperature. Heat to a temperature close to the temperature. For example, it is conceivable that the difference from the heat retention temperature is about 5 degrees or less. Alternatively, for example, the heating temperature by the temperature control mechanism is controlled to a temperature closer to the heat retention temperature than the room temperature.

The temperature is preferably the reaction temperature of the constant temperature nucleic acid amplification method that uses an appropriate reaction temperature. The appropriate reaction temperature is preferably in a range close to the optimum temperature of the enzyme. For example, it is in the range of about −5 ° C. to 0 ° C. More preferably, it is in the range of about −3 ° C. to 0 ° C. for the optimum temperature of the enzyme.
(3) A chemical analyzer including a structure in which a sample containing a biological material is introduced, a structure that holds a reagent to be reacted with the sample is contained, and includes a detection mechanism for the sample after reacting with the reagent, A driving mechanism for rotationally driving the structure and a mechanism for extracting a biological material from the sample, wherein at least one of the reagents is an enzyme-containing reagent, and the extracted biological material includes an enzyme-containing reagent. A container having a supply mechanism, a temperature control mechanism for controlling the temperature of the structure, a container in which the structure is stored, a tank in which the container is installed, a container for storing the tank, and an opening / closing mechanism And a first temperature control mechanism that is installed corresponding to the region in which the structure is accommodated, and that controls the temperature of the region where the extracted biological material of the structure is located, and the space in the tank Control the temperature of the fluid filled in the second Is a chemical analysis apparatus characterized by having degrees control mechanism.

The fluid filled in the tank internal space is a gas. For example, air may be used. From the viewpoint of suppressing oxidation or the like, a gas that suppresses oxidation of nitrogen or the like may be used.
(4) In the chemical analysis device of (3), the biological material is a nucleic acid, and the temperature of the biological material and the fluid in the tank is set before mixing the biological material and the reagent containing the enzyme. The chemical analyzer is controlled to a temperature closer to the predetermined temperature than the outside of the apparatus.
(5) In the chemical analysis apparatus of (3), the biological material is a nucleic acid, and the reagent temperature including the biological material and the enzyme is set to the temperature before mixing the biological material and the reagent including the enzyme. The chemical analyzer is controlled to a temperature closer to the predetermined temperature than the outside.
(6) In the chemical analysis device of (3), the biological material is a nucleic acid, and after mixing the biological material and the reagent containing the enzyme, a reaction solution of the biological material and the reagent containing the enzyme, The chemical analysis apparatus is characterized in that the temperature of the fluid in the tank is kept warm by operating at least the second temperature control means.

  If a temperature drop occurs when a reagent containing an enzyme is added to a sample, nucleic acid amplification due to the temperature drop will be affected. Thus, it is possible to construct a system that can have a stable amplification step that effectively suppresses deterioration of the reaction characteristics after mixing the sample and the reagent while suppressing deterioration of the characteristics of the reagent including the enzyme. it can.

  In addition, regarding the evaporation of the reaction liquid when controlling the temperature, it is possible to suppress the possibility that the reaction liquid evaporates and loses the amount of the problem as in the case of local heating as in the prior art, and is stable. Can be detected.

  According to the present invention, a chemical analyzer having improved reaction characteristics can be provided.

  A genetic test apparatus according to an embodiment of the present invention will be described in detail with reference to the drawings. It should be noted that the present invention is not intended to be limited to the contents disclosed in the present specification or claims, but is allowed to be modified based on other known techniques.

  FIG. 1 is a longitudinal sectional view showing a configuration of a genetic test apparatus according to an embodiment of the present invention. The genetic test apparatus 1 includes a holding disk 12 rotatably supported by a high-speed rotating motor 11 in a circular centrifuge tank 10, a plurality of test modules 2 arranged on the holding disk 12, and a perforation for controlling the flow of liquid. Machine 13, a test liquid temperature control device 16 for a module, a temperature control device 18 for a centrifuge tank with a built-in holding disk, and an amplification detection device 15. The inspection liquid temperature control device 16 is an apparatus that enables control of the inspection liquid temperature in the inspection port 390 and is configured by an electric heater. The centrifuge tank temperature control device 18 includes a heating electric heater 181, a circulation fan 182, and a temperature control device (PID control device in this embodiment) 183 that controls them.

  FIG. 2 is a view in which the centrifuge tank lid 9 in FIG. 1 is opened and the centrifuge tank 10 is viewed from above. It comprises a holding disk 12 that houses the inspection module 2 concentrically with the centrifuge tank 10. In order to arrange a large number of inspection modules 2 radially on the holding disk 12, the inner peripheral side surface of each inspection module 2 has a shape cut by a line inclined from the radial line of the holding disk 12. . Thereby, the inspection module 2 can be densely arranged on the holding disk 12, and the inspection efficiency is improved.

  The operator prepares the inspection module 2 for each inspection item and attaches it to the holding disk 12. FIG. 2 shows an example in which six test modules can be mounted and six specimens can be tested at the same time. Next, the genetic testing device 1 is activated.

  FIG. 3 is a configuration diagram of the inspection module 2. The inspection module 2 is configured by mounting the reagent cartridge 20 in which the transparent reagent cartridge cover 22 is joined to the reagent cartridge body 21 on the inspection cartridge 30 in which the transparent inspection cartridge cover 32 is joined to the inspection cartridge body 31. Each reagent is dispensed by a predetermined amount in each reagent container 220, 230, 240, 250, 260, 270, 280, 290 in advance.

Reagent outlets 221, 231, 241, 251, 261, and 271 that communicate with each reagent container are provided on the back surface of the reagent cartridge 20 shown in FIG. 4, and the reagent cartridge 20 shown in FIG. The test cartridge 30 is connected to each reagent inlet 321, 331, 341, 351, 361, 371. When the reagent cartridge 20 is mounted on the test cartridge 30, each reagent container communicates within the test cartridge through each reagent outlet and each corresponding reagent inlet.

  6 shows a main part of the longitudinal sectional view of the reagent cartridge 20 and the test cartridge 30 in the AA section shown in FIGS. 3 and 5, and FIG. 7 shows the above in a state where the reagent cartridge 20 is mounted on the test cartridge 30. The longitudinal cross-sectional view corresponding to AA part is shown. A reagent cartridge protective sheet 23 is adhered to the lower surface of the reagent cartridge 20 in order to prevent leakage and evaporation of the reagent stored in advance in the reagent cartridge 20, and the inner surface of the inspection cartridge 30 is attached to the upper surface of the inspection cartridge 30. In order to prevent contamination, the inspection cartridge protection sheet 33 is adhered.

  The operator peels off the reagent cartridge protection sheet 23 and the inspection cartridge protection sheet 33 and attaches the reagent cartridge 20 to the inspection cartridge 30. Projections (for example, 269 in FIG. 6) constituting the reagent outlets are fitted with the reagent inlets to position both cartridges, and the reagent does not leak out of the test cartridge. Alternatively, an adhesive may be attached to the inspection cartridge cover 32 to which the inspection cartridge protective sheet is adhered, thereby adhering the joint surface of the reagent cartridge to prevent the reagent from leaking.

  Note that a protrusion (for example, 269 in FIG. 6) provided on the reagent cartridge may be provided on the inspection cartridge side.

  The viral nucleic acid extraction and detection operations when whole blood is used as a sample will be described below with reference to FIGS.

  The operator injects whole blood collected by a vacuum blood collection tube or the like into the sample container 310 from the sample injection port 301 of the test cartridge 30, and peels off the reagent cartridge protection sheet 23 and the test cartridge protection sheet 33 shown in FIG. After that, the reagent cartridge 20 is mounted on the inspection cartridge 30 (FIG. 3). At this time, the sample inlet 301 of the test cartridge 30 is blocked by the reagent cartridge 20, so that the sample will not leak from the test cartridge 30 thereafter. Alternatively, a sample vent hole 313 is passed through the reagent cartridge 20 (FIGS. 2 and 3), and a filter that allows air to pass but does not allow the sample and its mist to pass is attached so that the sample can be vented when it flows. May be.

  When the necessary number of test modules 2 assembled in this manner are mounted on the holding disk 12 of FIG. 1 and the genetic test apparatus 1 is operated, viral genes are extracted from the whole blood, and the subsequent amplification process is performed. Finally, the gene is detected.

  Below, the flow state of the liquid in each operation | movement in the genetic test | inspection apparatus 1 is shown using FIG. 8 and FIG.

  After injecting whole blood 501 into the sample inlet 301, the holding disk 12 is rotated by the motor 11. The whole blood injected into the sample container 310 flows to the outer peripheral side by the action of the centrifugal force generated by the rotation of the holding disk 12, fills the blood cell storage container 311 and the serum quantitative container 312, and excess whole blood flows into the overflow tubule flow. It flows from the path 313 to the whole blood disposal container 315 through the overflow thick pipe channel 314. The whole blood disposal container 315 is provided with a whole blood disposal ventilation channel 318, and air can freely enter and exit from the reagent cartridge ventilation hole 202 through the test cartridge ventilation hole 302. Since the connection from the overflow narrow tube flow path 313 to the overflow large tube flow path 314 is abruptly expanded and is located on the innermost peripheral side (radius position 601) of the overflow thin tube flow path 313, whole blood flows into the overflow narrow tube flow path 313. It cuts at the connecting portion in a state where Accordingly, since no liquid can exist on the inner peripheral side from the radial position 601, the liquid level of the serum quantitative container 312 also becomes the radial position 601. The whole blood also flows into the serum capillary 316 branched from the serum quantification container 312, and the innermost peripheral portion of the whole blood is at the radial position 601 here.

  When the rotation is further continued, the whole blood 501 is separated into blood cells and serum or plasma (hereinafter referred to as “serum”) (centrifugation), the blood cells 502 move to the blood cell storage container 311 on the outer peripheral side, and the serum quantification container 312 has the serum 503. It becomes only.

  During the above series of serum separation operations, the vent holes 222, 232, 242, 252, 262, and 272 of each reagent container in the reagent cartridge 20 of FIG. 8 are covered with the reagent cartridge cover 22 (FIG. 5) so that the air flows. It is in a state that does not enter. Each reagent tries to flow out from the outer peripheral side of the reagent container due to the centrifugal force. However, since air does not enter the container, the pressure in the reagent container decreases, and the reagent cannot flow out in balance with the centrifugal force. However, when the rotational speed increases and the centrifugal force increases, the pressure in the reagent container gradually decreases, and bubbles are generated when the pressure is lower than the saturated vapor pressure of the reagent. Therefore, as shown in FIG. 9, the pressure drop in the reagent container is suppressed by adopting a flow path structure (return flow path, for example, 223) that once returns the reagent flowing out from the outer peripheral side of each reagent container to the inner peripheral side. And prevent the generation of bubbles. Thus, during the serum separation operation, each reagent does not flow while being held in the reagent container.

  When the serum separation operation is completed by rotating for a predetermined time, the test module 2 is stopped, and a part of the serum 503 in the serum quantitative container 312 flows into the capillary capillary 316 by surface tension, and the mixing unit 410 and the serum capillary 316 To the mixing part inlet 411, which fills the serum capillary 316.

  Thereafter, the perforator 13 makes holes one by one in the vent holes in the upstream portion of each reagent container, and the motor 11 is rotated to cause each reagent to flow by centrifugal force.

  The operation after serum separation is shown below. In the lysing solution container 220, a lysing solution 521 for dissolving a viral membrane protein in serum is dispensed. When the motor 11 is rotated after the perforator 13 has made a hole in the solution vent hole 222, the solution 521 passes from the solution container 220 through the solution return channel 223 by the action of centrifugal force, and the hygroscopic material 291 is removed. Then, it flows into the internal control container 290, and the solution flows into the mixing unit 410 while mixing with the internal control 590. The internal control is a nucleic acid or a synthetic product containing the nucleic acid, and is preferably lyophilized so that it can be stored for a long time. Therefore, when the solution flows into the internal control container 290, it mixes and flows out while dissolving the internal control 590.

  The hygroscopic material 291 is provided between the solution container 220 and the internal control container 290 so that the internal control 590 does not absorb the moisture of the solution 521. Hygroscopic materials include silica gel structures, porous structures using other materials or structures that make up fine flow paths such as fiber filters, and protrusions made of silicon or metal made by etching or machining. Use it.

  Further, since the innermost peripheral side of serum in the serum quantitative container 312 (radius position 601 at the end of serum separation) is located on the inner peripheral side from the mixing unit inlet 411 (radial position 602), the serum quantitative container is caused by a head difference due to centrifugal force. The serum in 312 and the serum capillary 316 flows into the mixing unit 410 from the mixing unit inlet 411 and is mixed in the mixing unit 410 with the dissolved solution in which the internal control flowing in at the same time is dissolved. The mixing unit 410 is composed of a member that mixes serum and lysate. For example, porous filters and fibers such as resin, glass and paper, or protrusions such as silicon and metal manufactured by etching or machining.

  Serum and lysate are mixed in the mixing unit 410 and flow into the reaction vessel 420. The reaction container 420 is provided with a reaction container ventilation channel 423, and air can freely enter and exit from the reagent cartridge ventilation hole 202 through the inspection cartridge ventilation hole 302. Since the branch 317 (radius position 603) from the serum quantitative container 312 to the serum capillary 316 is located on the inner peripheral side from the mixing section entrance 411 (radius position 602), all the serum in the serum capillary 316 is mixed by the siphon effect. Flows out. On the other hand, since the serum in the serum quantitative container 312 flows into the serum capillary 316 by centrifugal force, the serum flows out to the mixing part 410 until the liquid level of the serum in the serum quantitative container 312 reaches the branching portion 317 (radius position 603). Subsequently, when the serum level reaches the branching portion 317, air enters the serum capillary 316 and becomes empty, and the flow ends. That is, the serum in the serum quantitative container 312, the overflow capillary channel 313, and the serum capillary channel 316 from the radial position 601 to the radial position 603 at the end of serum separation flows out to the mixing unit 410 and mixes with the lysate.

  As described above, if the serum quantification container 312, the overflow capillary channel 313 and the serum capillary channel 316 from the radial position 601 to the radial position 603 are designed to have a predetermined volume (required serum volume), Even if the ratio is different for each whole blood sample, the serum used for the analysis can be quantified. For example, when the blood cell storage container has a volume of 250 microliters and the required serum volume is designed to be 200 microliters, if 500 microliters of whole blood is dispensed, 50 microliters of whole blood overflows into the whole blood disposal container 315. The remaining 450 microliters is separated into serum and blood cells, and 200 microliters of the separated serum flows out to the mixing unit 410. That is, for a whole blood sample of 450 microliters, a whole blood sample having a serum amount of 200 microliters or more can be analyzed with the device of this embodiment. For whole blood with a low serum ratio, the volume of the blood cell storage container may be increased to increase the number of whole blood samples.

  In the reaction container 420, the mixed serum and the lysate react. Since the liquid level in the reaction vessel 420 after the mixed solution of serum and lysate flows into the reaction vessel 420 is on the outer peripheral side of the innermost peripheral portion (radius position 604) of the reaction solution flow path 421, the reaction solution The innermost peripheral portion of the flow path 421 cannot be exceeded, and the mixed liquid is held in the reaction vessel 420 during rotation.

  The lysing solution functions to dissolve the membrane from viruses, bacteria, and the like in the serum to elute the nucleic acid, and further promotes adsorption of the nucleic acid to the nucleic acid binding member 301. Similarly, adsorption of the dissolved internal control 590 to the nucleic acid binding member 301 is promoted. As such a reagent, guanidine hydrochloride may be used for elution and adsorption of DNA, guanidine thiocyanate may be used for RNA, and a porous material such as quartz or glass or a fiber filter may be used as the nucleic acid binding member.

  After the serum and the lysate are held in the reaction container 420, the motor 11 is stopped, the additional liquid vent hole 232 for supplying air to the additional liquid container 230 is made by the punching machine 13, and the motor 11 is rotated again. Then, the additional liquid 531 flows from the additional liquid container 230 through the additional liquid return channel 233 into the reaction container 420 by the action of centrifugal force, and moves the liquid level of the mixed liquid in the reaction container to the inner peripheral side. When the liquid level reaches the innermost peripheral part (radius position 604) of the reaction liquid flow path 421, the mixed liquid flows out beyond the innermost peripheral part of the reaction liquid flow path, and passes through the converging flow path 701 to the nucleic acid binding member 301. Flows in. As the additional liquid, for example, the above-described dissolving liquid may be used.

  Depending on the sample, the wettability of the mixed liquid to the wall surface is good, and in the stopped state, the mixed liquid may flow in the reaction liquid channel 421 by capillary action. In this case, the additional liquid 531 is not required.

  When the mixture of the lysate and serum passes through the nucleic acid binding member in this manner, the target nucleic acid in the serum and the nucleic acid as the internal control are adsorbed to the nucleic acid binding member 301, and the liquid becomes a test port 390 that serves as an eluent recovery container. Flow into.

  The inspection port 390 is provided with an eluent recovery container vent channel 394, and air can freely enter and exit from the reagent cartridge vent 202 through the test cartridge vent 302. The waste liquid 391 after passing through the nucleic acid binding member 301 is once held in the eluent recovery container 390 for the waste liquid return channel 393 as in the case of the mixing container 420, but the eluent recovery container 390 is compared with the amount of waste liquid. Therefore, the waste liquid passes over the innermost peripheral side of the waste liquid return channel 393 and flows out to the waste liquid storage container 402 through the waste liquid outflow channel 399.

  Next, when the motor 11 is stopped and the perforator 13 opens a hole in the first cleaning liquid vent 242 for supplying air to the first cleaning liquid container 240, and then the motor 11 is rotated again, the centrifugal force acts. The first cleaning liquid flows from the first cleaning liquid container 240 into the nucleic acid binding member 301 via the first cleaning liquid return flow path 243 and the merge flow path 701 to wash unnecessary components such as proteins adhering to the nucleic acid binding member 301. As the first cleaning liquid, for example, the above-described dissolution liquid or a liquid with a reduced salt concentration may be used.

  The waste liquid after washing flows out to the waste liquid storage container 402 through the eluent recovery container 390, like the above-described mixed liquid.

  The same cleaning operation is repeated several times. For example, following the first cleaning liquid, with the motor stopped, the perforator 13 opens a hole in the second cleaning liquid vent hole 252 for supplying air to the second cleaning liquid container 250, and rotates the motor 11 again, so that the nucleic acid binding member Wash away unnecessary components such as salt adhering to 301. As the second cleaning liquid, for example, ethanol or an aqueous ethanol solution may be used.

  Similarly, a hole is made in the lid of the third cleaning liquid vent hole 262 for supplying air to the third cleaning liquid container 260. The third cleaning liquid flows directly into the eluent recovery container 390 and cleans components such as salt attached to the eluent recovery container 390. As the third cleaning liquid, for example, sterilized water or an aqueous solution whose pH is adjusted to 7 to 9 may be used.

  After washing the nucleic acid binding member 301 and the eluent recovery container 390 in this way, the process proceeds to the nucleic acid elution step.

  That is, with the motor stopped, a hole is made in the lid of the eluent vent hole 272 for supplying air to the eluent container 270 by the punching machine 13, the motor 11 is rotated again, and the eluent 571 flows through the nucleic acid binding member 301. . The eluent is a liquid that elutes nucleic acid from the nucleic acid binding member 301, and water or an aqueous solution adjusted to pH 7 to 9 may be used.

  After washing the nucleic acid binding member 301 and the eluent recovery container 390 in this way, the process proceeds to the nucleic acid elution step.

  That is, with the motor stopped, a hole is made in the eluent vent hole 272 for supplying air to the eluent container 270 by the perforator 13 and the motor 11 is rotated again to flow the eluent 571. The eluent is a liquid that elutes nucleic acid from the nucleic acid binding member 301, and water or an aqueous solution adjusted to pH 7 to 9 may be used. The amount of the liquid from which the nucleic acid has been eluted is smaller than the volume of the eluent recovery container 390, cannot exceed the innermost peripheral side of the waste liquid return channel 393, and is retained in the eluent recovery container.

  Next, the process proceeds to the nucleic acid amplification / detection step.

  FIG. 12 shows a test process when the Nucleic Acid Sequence-Based Amplification (NASBA) method, which is one of the constant temperature nucleic acid amplification methods, is used in this gene test apparatus. In the detection / amplification step, the nucleic acid is amplified by adding the amplification solution and the enzyme to the test solution containing the specimen sample in the test port 390 and holding the sample at a predetermined temperature for a predetermined time. At the same time, detection is performed by moving the detection optical cylinder 151 of the detection device 15 to a position where the test liquid in the test port 390 can be observed, and detecting the fluorescence emission amounts of the target nucleic acid and the internal control nucleic acid.

  Internal controls are pre-quantified nucleic acids or synthetic products containing nucleic acids, and are extracted, amplified, and detected using exactly the same reagents, cartridges, and testing equipment as the extraction, amplification, and detection of target nucleic acids in serum. Therefore, if the extraction / amplification / detection steps are functioning normally, a signal such as predetermined fluorescence or absorbance can be detected from the internal control. On the other hand, if the signal intensity is low or not detected at all, it can be seen that there is an abnormality in any of the extraction, amplification, and detection processes due to a defect in the reagent, cartridge, inspection device, or the like. Alternatively, the concentration of the target nucleic acid can be quantitatively evaluated by comparing the detection signal of the target nucleic acid with the detection signal of the internal control that has been quantified in advance.

  From the time when the extraction of the nucleic acid is finished, the temperature control of the centrifuge tank 10 is started and the control to the optimum enzyme temperature at which the air in the centrifuge tank 10 becomes the second reaction temperature, for example, 41 ° C. is started immediately. It does not matter if the temperature does not reach the optimum temperature.

  Next, since the extracted nucleic acid is contained in the inspection port 390, the amplification liquid 580 previously sealed in the inspection cartridge 30 is opened in a hole in the amplification liquid container 395, and the motor is rotated to rotate the amplification liquid. 580 enters inspection port 390. The amplification solution is a reagent for amplifying and detecting a nucleic acid, and preferably contains a fluorescent reagent in addition to deoxynucleoside triphosphate.

  Then, the test liquid temperature control device 16 is activated and control is started so that the test liquid reaches a denaturation temperature, for example, 65 ° C., which is the first reaction. After holding at 65 ° C. for about 5 minutes, the control is shifted to 41 ° C., which is the optimum enzyme temperature for the second reaction temperature. After holding at this temperature for 5 minutes, a hole is made in the enzyme container 396 and the holding disk 12 is rotated to put the enzyme 595 into the test port 390. Since the decrease in the liquid temperature immediately after the addition of the enzyme affects the subsequent amplification of the nucleic acid, it is necessary to prevent the temperature decrease as much as possible. Therefore, as described above, the temperature around the cartridge at the time of enzyme addition is increased in advance by the temperature control device in the centrifuge tank 10 so that the test solution and the reagent are maintained at the first reaction solution temperature of 41 ° C. It is mixed and there is no temperature drop.

  Since nucleic acid amplification proceeds in a steady state at 41 ° C. for 90 minutes, the amount of nucleic acid amplification can be detected by simultaneously detecting the amount of fluorescence. By using two types of fluorescent reagents having different wavelengths, it is possible to quantify in real time by comparing the amount of fluorescence emitted from the sample nucleic acid and the internal control.

  In addition, temperature control with high accuracy is possible in a short time by individual heating means by enabling the temperature control device of the inspection port to control the temperature to 65 ° C. Until the test cartridge is mounted, the manual operation is performed, but the subsequent steps from nucleic acid extraction to amplification can be fully automated.

  For this reason, the inspection port temperature control 16 and the centrifuge tank temperature control device 18 are provided in the embodiment of FIG.

  FIGS. 13 to 22 explain the liquid temperature control means in the inspection port, and FIG. 13 shows the holding disk 12 and the AA cross-sectional view of the inspection cartridge 2 in the embodiment of FIGS.

  The embodiment shown in FIG. 14 uses the inspection cartridge 2 provided with a heater 162 for controlling the temperature of the inspection liquid 550, and is energized by a power supply line (not shown) to generate heat. By providing the cartridge with a heating means, it is possible to prevent variation due to contact thermal resistance between the heating unit and the outer wall of the inspection port 390, and to perform stable temperature control. In this case, in order to measure the fluorescence emission amount in the inspection port 390 with the detection optical cylinder 151, it is necessary to provide holes 162b and 12b in the heater 162 and the holding disk, respectively, through which the inside of the inspection port 390 can be optically observed.

  Further, FIG. 15 shows another example of the present embodiment. In this embodiment, a heater 162 serving as a heating means is built in the rotating holding disk 2 side. With such a configuration, the heating unit can be separated from the cartridge side, so that the disposable test cartridge 2 can be inexpensively configured.

The embodiment of FIG. 16 shows yet another example. In this embodiment, the Peltier element 164 is used for temperature control of the test liquid 550. The outer wall of the inspection port 390 containing the inspection liquid 550 and the heat block 163 are thermally connected. The amount of heating can be controlled by changing the magnitude of the applied current to the Peltier element 164, and not only the temperature can be raised but also lowered by changing the forward / reverse direction. Thereby, when the temperature drops from 65 ° C. to 41 ° C., the temperature can be controlled for a short time. Note that the embodiment of FIG. 17 in which 164b serves as the heat absorption part of the Peltier element shows still another example, and shows a state in which the temperature monitor element 165 is placed in the test liquid 550. By directly placing a thermocouple, thermistor, and platinum resistor coated with a substance that does not inhibit gene amplification on the surface of the temperature sensing portion into the test solution 550, the test solution temperature can be measured and controlled with high accuracy. Further, the object of the present embodiment can be achieved even if the inspection apparatus main body is provided with a cleaning device for the temperature sensor element 165 and measurement and cleaning are repeated for each inspection.

  The embodiment of FIG. 18 shows still another example and shows a configuration in which the temperature sensor 165 is built in a heat block 163 that encloses a test liquid container. With such a configuration, the temperature can be measured without replacing the temperature sensor 165, and an inexpensive structure can be realized.

  The embodiment of FIG. 19 shows another example, and the surface temperature of the cartridge 2 is measured using an infrared radiation thermometer 166. With such a configuration, liquid measurement can be performed without contact with the rotating holding disk 12. In this case, it is necessary to measure and grasp the difference between the test solution temperature and the cartridge surface temperature in advance.

  The embodiment of FIG. 20 shows still another example, and the temperature of the heat block 163 wrapping the inspection port 390 is monitored using the same infrared radiation thermometer 166 as in FIG. With this configuration, since the emissivity can be defined by applying a black body paint or the like having a high emissivity to the heat block 163 in advance, it is possible to perform measurement with relatively high accuracy in infrared temperature measurement. In the embodiment of FIGS. 19 and 20, an infrared radiation thermometer is shown as an example. However, even if the temperature measurement method is such that a temperature-sensitive liquid crystal is applied to the cartridge and an image taken with a CCD camera or the like is processed, Measurement is possible.

  In the embodiment of FIG. 21, yet another example is shown. An infrared lamp 167 is used as a heating method. Also in this case, heating without contact with the rotating holding disk 12 is possible. During the infrared lamp irradiation, the fluorescence detection is not affected by stopping the lamp heating in the fluorescence measurement performed by the optical detection cylinder 151.

  In the embodiment of FIG. 22, yet another example is shown. Electromagnetic conduction is used as a heating method. A metal heat block 163 having a predetermined electrical resistance is arranged around the inspection port 390. By energizing the coil 169 connected to the AC generation source 168, an eddy current flows in the heat block 163 by electromagnetic induction, and heating can be performed by Joule heat generation. The amount of heating can be varied depending on the strength of the alternating current and the distance between the coil 169 and the heat block 163, and heating control without contact with the holding disk 12 becomes possible. In this case, it is desirable that the holding disk 12 be made of a non-energized material. The effect of this embodiment can also be achieved by a method in which the coil 169 is replaced with a microwave oscillation device (magnetron) in the embodiment of FIG. 22 to induce vibration of water molecules in the test liquid 550 and directly heat it.

  Next, FIG. 23 shows a temperature change of the water vapor partial pressure of the humid air under a general atmospheric pressure. The inside of the test port 390 is considered to have a relative humidity of 100% because an evaporation phenomenon occurs at 41 ° C., the optimal enzyme temperature, due to control during the nucleic acid amplification process. Therefore, evaporation, which is a diffusion phenomenon using a difference in water vapor partial pressure from the air vent hole opened to the inspection cartridge 2 to the ambient air as a drive source, is likely to occur. When evaporation occurs, it becomes difficult to detect the amount of fluorescent light emission due to a lack of the test solution, and temperature distribution in the test solution is likely to occur due to temperature equilibrium at the gas-liquid interface of the test solution. Further, the concentration of the inspection liquid changes due to condensation of the evaporated inspection liquid vapor on the inner surface of the inspection cartridge cover 32.

  Therefore, as shown in FIG. 23, during the optimum temperature control, the difference in water vapor partial pressure can be reduced and the evaporation of the inspection liquid 550 can be prevented as the air temperature around the inspection cartridge 2 is increased as compared with the room temperature. Therefore, as shown in FIG. 1, the heating control means in the centrifuge tank 10 is provided.

  By controlling the inside of the centrifuge tank to a temperature close to the second reaction solution temperature, as described above, it is possible to prevent the deactivation of the enzyme and the evaporation of the test solution from the test module. Further, the temperature distribution is small and the concentration distribution can be reduced by controlling the temperature of the test liquid from the surroundings. Furthermore, the measurement error due to the displacement of the test solution temperature can be reduced.

  Next, temperature control of the air in the centrifuge tank 10 will be described.

  In the embodiment of FIG. 1, a film-like rubber heater 181 is used as the heating means of the centrifuge tank 10. The metal portion of the centrifuge tank is used as a heat transfer area, and the heat exchange efficiency is enhanced by a wide area. The heater may be coiled and wound around the outer periphery or inner periphery of the centrifuge tank.

  In the embodiment of FIG. 24, a fixed constant temperature disk 197 is incorporated in addition to the example of FIG. The holding disk 12 is installed with a predetermined gap. When the constant temperature disk 197 is heated to a predetermined temperature by a method such as a heater (not shown) and the rotatable holding disk 12 is rotated, convection is generated in the air in the centrifuge tank 10 to promote heat transfer, and the temperature is maintained from the constant temperature disk. Heat is transferred to the disk 12. With such a configuration, temperature control is quickly performed by the rotating operation of the disk. In addition, if the temperature of both disks is reversed and the temperature is set to a low temperature such as circulating cold water in the constant temperature disk, the temperature of the holding disk can be lowered in a short time.

  In the embodiment of FIG. 25, as a temperature control means in the centrifuge tank 10, an air passage 184 is provided outside the centrifuge tank 10, the air in the centrifuge tank 10 is flowed, heated by the heater 185, and repeatedly circulated. This is the structure. With such a configuration, the air velocity in the vicinity of the heater can be increased, forced convection heat transfer is enhanced, and heat exchange efficiency is high, so the heater capacity may be small.

  In the embodiment of FIG. 26, another example is shown, and a humidifying function by a humidifier 186 is added to the structure of FIG. With such a configuration, the humidity in the centrifuge tank can be increased, and the evaporation of the test solution can be further prevented as shown by the characteristics of the water vapor partial pressure in FIG.

  The embodiment of FIG. 27 shows yet another example. As a heating means of the centrifuge tank 10, a hot water circulation system is adopted. A hot water coil 191 capable of flowing hot water is wound inside the centrifuge tank 10 inside. By circulating the hot water maintained at a constant temperature in the coil 191 by the constant temperature water tank 188 by the operation of the pump 189, the inside of the centrifugal tank 10 can be stabilized at a constant temperature with high accuracy.

  FIG. 28 shows a case where the centrifuge tank 10 is heated by the refrigeration cycle. As components, a condenser pipe 192a and an evaporator coil 192b attached to the centrifuge tank 10, and a compressor, a four-way valve, and an expansion valve constitute a refrigeration cycle. The centrifuge tank 10 can be heated with the high-temperature superheated gas discharged from the compressor and the condensate by rotating the compressor with the chlorofluorocarbon inside and rotating the expansion valve to an appropriate degree of opening. By switching the flow direction of the refrigerant by switching the four-way valve, it is possible to make 192a a low temperature evaporation coil and 192b a high temperature condensation coil. In this case, the temperature can be freely changed by a combination of the compressor rotational speed and the expansion valve opening.

  The embodiment of FIG. 29 has a structure provided with an upper blower 196 of the centrifuge tank lid 9 in addition to the embodiment of FIG. With such a configuration, the circulation of air inside can be promoted and the temperature distribution of air in the centrifuge tank 10 can be reduced. In FIG. 29, the blower 196 is shown as being exposed in the centrifuge tank 10, but by providing a cover, turbulence due to air convection generated by high-speed rotation of the holding disk 12 can be prevented. In addition, it is preferable to retain the heat of the heater 181 effectively.

  FIG. 30 shows another example of the present embodiment. In the nucleic acid detection process of the inspection process, an operation of rotating the holding disk 12 at a high speed is added immediately before the enzyme is added. By such an operation, frictional heat is generated between the air in the centrifuge tank 10 and the holding disk 12, and the air temperature in the centrifuge tank 10 can be increased in a short time. Further, the air inside the centrifuge tank 10 is mixed by the rotating operation of the holding disk, and the temperature can be made uniform.

  FIG. 31 is a diagram illustrating temporal changes in ON / OFF of the temperature control of the inspection port 390 and the temperature control of the centrifuge tank 10. (A) shows the temperature control of the inspection port. The case where control is started at 65 ° C. as an example of the first optimum temperature simultaneously with the end of the extraction step, and is controlled to 41 ° C. as an example of the second optimum temperature after about 5 to 10 minutes after that. Under the conditions of (a), (b) to (e) show ON / OFF states of the temperature control of the centrifuge tank 10. First, (b) starts temperature control of the centrifuge tank 10 simultaneously with the end of the extraction process. (C) starts control in the middle of the extraction process. (D) is performed in the middle of the detection process after the extraction process is completed. (E) ends in the middle of the detection process. In this control, it is possible to prevent the influence due to the high temperature in the extraction process in the next inspection module.

  Next, FIG. 32 shows the time change of the test solution temperature according to the temperature control result of FIG. FIG. 32 (a) corresponds to (c) in FIG. 31, and the control is started from the middle of the extraction process, and reaches the second reaction temperature of 41 ° C. at the end of the extraction process. This temperature control has an effect of reducing the evaporation amount of the reaction liquid during the control of the first reaction temperature of 65 ° C. In FIG. 32 (b), the second reaction temperature is reached in the middle of the first reaction temperature control. In (c), it is achieved according to the second reaction temperature. This temperature control has the effect of minimizing the influence of inactivation due to the temperature of the enzyme to be added.

  FIG. 33 shows the time trend of the temperature rise of the test solution. In FIG. 33, the temperature control of the centrifuge tank is started simultaneously from the start of extraction, but in (a), (b), and (c), the time until the temperature becomes constant is compared and shown. In (a), the second reaction liquid temperature is reached between the extraction and detection steps. In this case, since the water vapor partial pressure difference is small during the first reaction temperature control, the evaporation amount of the test solution can be minimized. In (b), it is achieved in the middle of the control of the first reaction liquid temperature. Further, in (c), the temperature has reached the detection start time of the second reaction liquid temperature. In the case of (c), the time at a relatively high temperature is short and the influence of enzyme deactivation can be minimized.

  In the present embodiment, the optimum enzyme temperature is shown, but there is no problem as long as the temperature is within the temperature range where the enzyme reacts. For example, even if the lower limit is in the range from the optimum temperature -5 ° C to the optimum temperature, etc. The effect of form is achieved.

  Further, it may be a testing apparatus that can be operated by inputting in advance a temperature control range that takes into account the optimum temperature of the enzyme for each test target or reagent to be used. In this case, the inspection time can be shortened.

The present embodiment described so far has the following forms.
(1) A chemical analyzer including a structure in which a sample containing a biological material is introduced, a structure that holds a reagent to be reacted with the sample is accommodated, and includes a detection mechanism for the sample after reacting with the reagent, The fluid surrounding the structure is controlled to an appropriate reaction temperature of the reaction reagent after extraction of the biological material in the sample from the structure or during the extraction. Specifically, at least one of the reagents is a reagent containing an enzyme, a mechanism for extracting a biological material from the sample, a mechanism for supplying a reagent containing the enzyme to the extracted biological material, and the structure A temperature control mechanism for controlling the temperature of the biological material, and after the step of extracting the biological material is started and before the reagent containing the enzyme is reacted with the biological material, It is controlled to raise the temperature of the reagent containing the enzyme. As the structure, the inspection difference module 2 (FIGS. 2 and 3) can be used.

  It is characterized in that the fluid surrounding the structure is controlled to an appropriate reaction temperature of the reaction reagent after extraction of the biological material in the sample with the structure or during the extraction.

The biological material can be, for example, the nucleic acid described in the examples. Or it can be DNA, RNA, or protein.
(2) or in the chemical analyzer of (1), the biological material supplied with the enzyme is kept at a predetermined temperature for a predetermined time, and the heated biological material is controlled to be detected by the detection mechanism, In the step of raising the temperature, the reagent containing the enzyme is controlled so as to approach the heat retention temperature.

  For example, as illustrated in the embodiments, the predetermined temperature is a heat retention temperature such as an optimum temperature. Generally, the temperature is higher than room temperature. Heat to a temperature close to the temperature. For example, it is conceivable that the difference from the heat retention temperature is about 5 degrees or less. Alternatively, for example, the heating temperature by the temperature control mechanism is controlled to a temperature closer to the heat retention temperature than the room temperature. Here, room temperature is assumed to be about 10-30 ° C.

The temperature is preferably the reaction temperature of the constant temperature nucleic acid amplification method that uses an appropriate reaction temperature. The appropriate reaction temperature is preferably in a range close to the optimum temperature of the enzyme. For example, it is in the range of about −5 ° C. to 0 ° C. More preferably, it is in the range of about −3 ° C. to 0 ° C. for the optimum temperature of the enzyme.
(3) or a chemical analyzer, comprising a drive mechanism for rotationally driving the structure and a mechanism for extracting a biological substance from the sample, wherein at least one of the reagents is a reagent containing an enzyme A mechanism for supplying an enzyme to the extracted biological material; a temperature control mechanism for controlling the temperature of the structure; a storage part for the structure; a tank in which the storage part is installed; and the tank A first container for controlling a temperature of a region where the extracted biological material of the structure is located, the container being provided corresponding to a region in which the structure is accommodated. A temperature control mechanism and a second temperature control mechanism for controlling the temperature of the fluid filled in the space in the tank are provided.

  The biological material in the structure and the reagent used for the reaction are mixed by the action of centrifugal force.

  The first temperature control mechanism can be provided in a rotating disk portion that is a housing portion for the structure. Thereby, temperature control of a specific area | region can be performed among the flow paths of a structure. Or it can also arrange | position through the space | interval with the said disk facing the test | inspection module 2 of a accommodating part.

The fluid filled in the tank internal space is a gas. For example, air may be used. From the viewpoint of suppressing oxidation or the like, a gas that suppresses oxidation of nitrogen or the like may be used.
(4) or the second temperature control mechanism in the chemical analysis apparatus of (3), before starting the extraction step of the biological material and before supplying the reagent containing the enzyme to the biological material. And a control for raising the temperature in the tank by operating the apparatus.

In addition, for example, the extraction step is within the range of the step of removing the target nucleic acid from the start of the step of centrifuging from the biological sample.
(5) Or, in the chemical analyzer of (3), the first temperature control mechanism between the start of the biological substance extraction step and the supply of the reagent containing the enzyme to the biological substance. And a control for operating the second temperature control mechanism to raise the temperature in the tank.
(6) Or, in the chemical analyzer of (3), the second temperature control mechanism has a stirring mechanism for stirring the gas in the tank.
(7) In the chemical analysis device of (3), the biological material is a nucleic acid, and before mixing the biological material and the reagent containing the enzyme, A: the biological material and the fluid in the tank The temperature is controlled to a temperature closer to the predetermined temperature than the outside of the device, or B: the temperature of the reagent containing the biological material and the enzyme is controlled to a temperature closer to the predetermined temperature than the outside of the device.

  By doing in this way, the temperature fall at the time of mixing with the test solution containing an enzyme and sample nucleic acid can be prevented, and the effect which prevents enzyme deactivation is acquired.

  Or C: after mixing the biological material and the reagent containing the enzyme, at least the second temperature control of the reaction solution of the biological material and the reagent containing the enzyme and the temperature of the fluid in the tank The temperature is controlled by operating the means. By doing in this way, since the difference of the water vapor partial pressure in a test | inspection port and a centrifuge tank can be made small, evaporation of a test | inspection liquid can be suppressed. Further, the temperature of the test solution can be made uniform, and the concentration distribution due to evaporation and condensation of the test solution during amplification and detection can be made uniform. Furthermore, since the temperature distribution of the test solution is reduced, the temperature measurement accuracy at the temperature control position can be improved.

  In order to control the fluid to the reaction temperature, at least one of an electric heater, hot water circulation, and a condenser of a refrigeration cycle is used.

  In the PCR amplification method, as described above, temperature control is repeated between predetermined temperatures, but it is not necessary to dispense reagents in the middle of the cycle. On the other hand, in the Nucleic Acid Sequence-Based Amplification (NASBA) method of the constant temperature nucleic acid amplification method, it is necessary to add an enzyme under a constant temperature condition.

It is a longitudinal cross-sectional view of the genetic testing apparatus of one Example of this invention. It is an upper arrow figure of the centrifuge tank of FIG. It is an external view of an inspection cartridge. It is a back external appearance figure of the non-mounting of a reagent cartridge. It is an external view of a test cartridge without a reagent cartridge. It is sectional drawing at the time of the non-connection of a test | inspection cartridge and a reagent cartridge. It is sectional drawing at the time of a connection of a test | inspection cartridge and a reagent cartridge. It is an upper arrow figure of a reagent cartridge. It is an upper arrow view of a test | inspection cartridge. It is an external view of an inspection module integrated with a reagent cartridge. It is an upper arrow figure of the inspection module of a reagent cartridge integrated type. It is a flowchart which shows an example of an inspection process. It is a figure which shows the cross section explaining the temperature control means of a test liquid. It is a longitudinal cross-sectional view explaining the test liquid temperature control means of a test cartridge. It is a longitudinal cross-sectional view explaining the test liquid temperature control means of a test cartridge. It is a longitudinal cross-sectional view explaining the test liquid temperature control means of a test cartridge. It is a longitudinal cross-sectional view explaining the test liquid temperature control means of a test cartridge. It is a longitudinal cross-sectional view explaining the test liquid temperature control means of a test cartridge. It is a longitudinal cross-sectional view explaining the test liquid temperature control means of a test cartridge. It is a longitudinal cross-sectional view explaining the test liquid temperature control means of a test cartridge. It is a longitudinal cross-sectional view explaining the test liquid temperature control means of a test cartridge. It is a longitudinal cross-sectional view explaining the test liquid temperature control means of a test cartridge. It is a figure of the water vapor partial pressure under atmospheric pressure explaining the effect of this invention. It is a longitudinal cross-sectional view of the test | inspection apparatus explaining the Example of the temperature control of the centrifuge tank of this invention. It is a longitudinal cross-sectional view of the test | inspection apparatus explaining the Example of the temperature control of the centrifuge tank of this invention. It is a longitudinal cross-sectional view of the test | inspection apparatus explaining the Example of the temperature control of the centrifuge tank of this invention. It is a longitudinal cross-sectional view of the test | inspection apparatus explaining the Example of the temperature control of the centrifuge tank of this invention. It is a longitudinal cross-sectional view of the test | inspection apparatus explaining the Example of the temperature control of the centrifuge tank of this invention. It is a longitudinal cross-sectional view of the test | inspection apparatus explaining the Example of the temperature control of the centrifuge tank of this invention. It is a longitudinal cross-sectional view of the test | inspection apparatus explaining the Example of the temperature control of the centrifuge tank of this invention. It is a temperature control timing diagram of a test solution and a centrifuge tank. It is a temperature control timing diagram of a test solution and a centrifuge tank. It is a temperature control timing diagram of a test solution and a centrifuge tank.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Gene test apparatus, 2 ... Test module, 10 ... Centrifugal tank, 12 ... Holding disk, 15 ... Detection mechanism, 16 ... Test port temperature control apparatus, 18 ... Centrifugal tank temperature control apparatus, 20 ... Test cartridge, 30 ... Reagent cartridge.

Claims (10)

  A chemical analysis apparatus including a structure in which a sample containing a biological material is introduced, a structure holding a reagent to be reacted with the sample, and having a structure for detecting the sample after reacting with the reagent, comprising: One is a reagent containing an enzyme, and includes a mechanism for extracting a biological substance from the sample, a mechanism for supplying a reagent containing the enzyme to the extracted biological material, and a temperature control mechanism for controlling the temperature of the structure. The temperature control mechanism controls the temperature of the reagent containing the enzyme after the step of extracting the biological material is started and before the reagent containing the enzyme is reacted. A chemical analyzer characterized by   2. The chemical analyzer according to claim 1, wherein the biological material supplied with the reagent containing the enzyme is amplified by keeping the temperature at a predetermined temperature for a predetermined time, and the detection mechanism detects the biological material after the temperature is maintained. The chemical analysis apparatus is characterized in that in the step of raising the temperature, the reagent containing the enzyme is controlled so as to approach the heat retention temperature.   A chemical analysis apparatus including a structure in which a sample containing a biological material is introduced, a structure holding a reagent to be reacted with the sample, and a reaction mechanism for the sample after reacting with the reagent is provided. A mechanism for rotating and driving, and a mechanism for extracting biological material from the sample, wherein at least one of the reagents is a reagent containing an enzyme, and a mechanism for supplying the extracted biological material with the reagent containing the enzyme And a temperature control mechanism for controlling the temperature of the structure, and a storage section of the structure, a tank in which the storage section is installed, and a container that stores the tank and includes an opening / closing mechanism. A first temperature control mechanism that is installed corresponding to the region in which the structure is accommodated and controls the temperature of the region where the extracted biological material of the structure is located; and the space in the tank is filled Second temperature control to control the temperature of the fluid Chemical analysis apparatus characterized by having mechanisms.   4. The chemical analysis apparatus according to claim 3, wherein the second temperature control mechanism is operated to start the extraction of the biological material and before supplying the reagent containing the enzyme to the biological material. A chemical analyzer comprising a control for raising the temperature inside.   4. The chemical analyzer according to claim 3, wherein the first temperature control mechanism and the second temperature are between the start of the biological material extraction step and the supply of the reagent containing the enzyme to the biological material. A chemical analyzer comprising a control for operating a control mechanism to raise the temperature in the tank.   4. The chemical analyzer according to claim 3, wherein the second temperature control mechanism has a stirring mechanism for stirring the gas in the tank.   4. The chemical analysis apparatus according to claim 3, wherein the biological material is a nucleic acid, and the temperature of the biological material and the fluid in the tank is set to the outside of the device before mixing the biological material and the reagent containing the enzyme. The chemical analyzer is controlled to a temperature closer to the predetermined temperature.   4. The chemical analyzer according to claim 3, wherein the biological material is a nucleic acid, and the reagent temperature containing the biological material and the enzyme is set to be higher than that outside the device before mixing the biological material and the reagent containing the enzyme. A chemical analyzer that controls the temperature to be close to the predetermined temperature.   4. The chemical analyzer according to claim 3, wherein the biological material is a nucleic acid, and after mixing the biological material and a reagent containing the enzyme, a reaction solution of the biological material and the reagent containing the enzyme and the inside of the tank A chemical analysis apparatus characterized in that the temperature of a fluid is controlled to keep the temperature by operating at least the second temperature control means.   4. The chemical analyzer according to claim 3, wherein the biological material is a nucleic acid.

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