JP6675168B2 - Nucleic acid testing equipment - Google Patents

Description

  An embodiment of the present invention relates to a nucleic acid testing device.

  As this type of inspection device, one using a current detection type DNA chip is known. This current detection type DNA chip arranges at least one DNA probe having a known base sequence on a plurality of electrodes provided on a substrate, and when the DNA probe and DNA contained in a test object are bonded, It detects the flowing current and specifies the type of DNA contained in the test object.

  However, in the conventional current detection type DNA chip, the number of electrodes provided on the substrate was small, and the number of detectable genes was small.

  In addition, before performing the above-described DNA test by current detection, the DNA contained in the test target must be amplified. Conventionally, after nucleic acid amplification is performed in another container, the DNA test is performed by relocating the container, which is labor-intensive for an operator, and there is a problem that the DNA test cannot be performed in a short time.

Japanese Patent No. 4729030

  The problem to be solved by the present invention is to provide a nucleic acid testing device capable of performing a nucleic acid test in a short time and with high accuracy without bothering an operator.

In the nucleic acid detection device attached to the nucleic acid testing device according to the present embodiment, the storage unit stores at least the sample. The amplification unit amplifies the nucleic acid contained in the specimen sample stored in the storage unit. The first flow path moves the specimen sample from the storage unit to the amplification unit. The detection unit detects a nucleic acid contained in the specimen sample that has undergone nucleic acid amplification in the amplification unit. The second flow path moves the sample from the amplification unit to the detection unit.
In the nucleic acid testing device, the first opening / closing unit opens / closes the first flow path. The second opening and closing unit opens and closes the second flow path. The heating unit heats the amplification unit. The control unit controls the first and second opening and closing units to open and close in a predetermined order, and controls the heating unit to heat the amplification unit in conjunction with the opening and closing operations of the first and second opening and closing units.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is an external view of a nucleic acid testing system including a nucleic acid testing device according to one embodiment, a unit configuration diagram of the nucleic acid testing device, and a diagram showing opening and closing of a tray provided in the nucleic acid testing device. FIG. 4 is a diagram illustrating an example of a GUI screen displayed on a display screen of the information processing device. FIGS. 3A to 3C are diagrams for explaining that a DNA probe and a DNA to be tested are complementary. FIGS. 4A to 4D are diagrams illustrating a base sequence detection chip. FIGS. 5A and 5B are waveform diagrams showing an example of a measurement result of an oxidation current detected from a base sequence detection chip. FIG. 2 is an external view of a nucleic acid test card. FIG. 3 is a perspective view showing a configuration of a nucleic acid test card. FIG. 3 is a perspective view showing a configuration of four syringes. The figure which shows the flow-path model of a nucleic acid test card. Sectional drawing of a valve NOV. The figure which shows the structure of an NC valve. The figure which shows the flow-path model of a nucleic acid test card. FIG. 2 is a plan view showing a detailed structure of an amplification channel. The top view which shows the detailed structure of a DNA chip. FIG. 2 is a block diagram of a control system of the nucleic acid testing device according to one embodiment. The figure of the air flow path by the housing fan in a nucleic acid testing device. FIG. 2 is an external view of a heater and a Peltier element. FIG. 5 is a plan view showing an arrangement of five motors and a syringe rod, an NCV rod, a NOV rod, a temperature control support, and a probe support driven by the motors. Sectional drawing of a rack gear. FIG. 2 is a plan view showing a syringe rod driven by a syringe shaft motor. The perspective view of a punch. FIGS. 22A and 22B show examples in which the width of the punch is changed and pressed against a syringe. The figure which shows the relationship between the position shift of a syringe, and the liquid sending amount from a syringe. The figure which made the syringe rod the taper structure. FIG. 2 is a plan view showing an NCV rod driven by an NCV axis motor. (A), (b) and (c) are figures which show the tip shape of a fork part. FIG. 4 is a plan view showing a NOV rod driven by a NOV axis motor. (A) and (b) are the top view and perspective view which show the temperature control support body driven by the heater Peltier shaft motor. (A) and (b) are a plan view and a perspective view showing a probe support driven by a probe shaft motor. The figure which shows the structure of the pin for positioning. 5 is a flowchart illustrating an example of a drive sequence of each motor by a control unit. The figure which shows the movement state of the liquid on a nucleic acid test card.

  Hereinafter, an embodiment of the present invention will be described with reference to the drawings. In the following embodiments, the characteristic configuration and operation in the nucleic acid testing device will be mainly described, but the nucleic acid testing device may have configurations and operations omitted in the following description. However, these omitted configurations and operations are also included in the scope of the present embodiment.

(overall structure)
FIG. 1A is an external view of a nucleic acid testing system provided with a nucleic acid testing device according to the present embodiment, FIG. 1B is a unit configuration diagram of the nucleic acid testing device, and FIG. 1C is a tray provided in the nucleic acid testing device. FIG. As shown in FIG. 1A, the nucleic acid testing system 100 includes a nucleic acid testing device 110 and an information processing device 150. The nucleic acid testing device 110 is for testing a nucleic acid contained in a specimen sample, as described later. The information processing device 150 instructs the nucleic acid testing device 110 for testing conditions and the start of testing, and performs analysis and display of test results by the nucleic acid testing device 110.

  The nucleic acid testing device 110 according to the present embodiment is characterized in that a plurality of sample samples can be tested at once. In the example of FIG. 1B, four test units 1, 2, 3, and 4 and a control board 15 are provided so that four types of sample samples can be tested at a time. Each of the test units 1, 2, 3, and 4 operates independently and independently under the control of the control board 15, and each of the test units 1, 2, 3, and 4 performs a nucleic acid test on a separate sample sample at a different timing. be able to.

  As will be described later, individual sample samples are put into and taken out of the nucleic acid testing apparatus 110 through the opening / closing ports 101, 102, 103, and 104 in a state of being inserted into a removable nucleic acid testing card (nucleic acid detecting device) 700. . In order to facilitate taking in and out, a tray 114 (mounting unit) as shown in FIG. 1C is provided for each of the inspection units 1, 2, 3, and 4, and these trays 111, 112, 113, and 114 are provided. After placing the nucleic acid test cards 700, the nucleic acid amplification and the nucleic acid test can be automatically performed thereafter.

  The nucleic acid testing apparatus 110 according to the present embodiment does not have a setting input function and a display function by itself. Therefore, it is possible to reduce the size and cost of the nucleic acid testing device 110. The setting input to the nucleic acid test device 110 and the analysis of the nucleic acid test result are performed by the information processing device 150 connected to the nucleic acid test device 110. Since the information processing device 150 can be constituted by a general-purpose computer such as a commercially available PC, it can be introduced at a low cost, and the maintenance cost is not so high. The nucleic acid testing device 110 and the information processing device 150 transmit and receive various information via a general-purpose communication interface such as a USB (Universal Serial Bus). As described above, by constructing the nucleic acid testing system with the nucleic acid testing device 110 and the information processing device 150, the maintenance management of the nucleic acid testing device 110 is facilitated, and the setting input to the nucleic acid testing device 110 and the analysis of the test result are performed. And make it easier to display.

  FIG. 2 is a diagram illustrating an example of a GUI screen displayed on the display screen of the information processing device 150. On the GUI screens 201, 202, 203, and 204 of FIG. 2, setting input items of four inspection units are displayed, respectively. According to the GUI screens 201, 202, 203, and 204, various types of information on each sample sample can be input for each of the test units 1, 2, 3, and 4. The various types of input information are transmitted from the information processing device 150 to the nucleic acid testing device 110 as necessary, and are set in the nucleic acid testing device 110. For example, in the GUI screens 201, 202, 203, and 204, selection buttons 211, 212, 213, and 214 that are selected when performing a nucleic acid test using each of the test units 1, 2, 3, and 4 are displayed. Test start buttons 221, 222, 223, and 224 for instructing start and display areas 231, 232, 233, and 234 for displaying the progress of the test in the nucleic acid test apparatus 110 are provided. The information processing device 150 and the nucleic acid testing device 110 transmit and receive various types of information by wire or wirelessly, and information obtained by selecting the selection buttons 211, 212, 213, 214 and the test start buttons 221, 222, 223, 224 is The information is transmitted to the nucleic acid testing device 110 instantaneously, and the test progress information in the nucleic acid testing device 110 is periodically transmitted from the nucleic acid testing device 110 to the information processing device 150.

  The GUI screens 201, 202, 203, and 204 displayed on the display screen of the information processing device 150 can be arbitrarily changed by software. Therefore, the GUI screens 201, 202, 203, and 204 in FIG. 2 are only examples. By updating the software, a GUI screen that is easy for the user to use can be provided, and a new type of nucleic acid test can be easily handled.

(Basic principle of nucleic acid test)
Before describing the nucleic acid testing apparatus 110 according to the present embodiment in detail, the basic principle of the nucleic acid testing employed in the present embodiment will be described.

  DNA has a double-stranded structure in which two chains each having a sequence of four bases of A (adenine), T (thymine), G (guanine), and C (cytosine) are connected. In a double-stranded structure, they bind with a specific combination of base A and base T, and base G and base C. Since DNA fragments can be easily synthesized, in the DNA chip 500, a single-stranded base sequence whose sequence is known is immobilized on the electrode as a probe. The DNA of the sample is also made single-stranded, and is reacted with the DNA probe immobilized on the electrode. If the sequence of the DNA in the sample is complementary to the sequence of the DNA probe, it will bind and become double-stranded. For example, as shown in FIG. 3 (a), when the DNA probes are arranged in the order of TAGAC and the DNA of the sample is arranged in the order of ATCTG (FIG. 3 (b)), the sequences are mutually different. Since they are complementary, these single-stranded DNAs shown in FIG. The hybridization of the base sequence of the sample with the base sequence of the complementary DNA probe to form a double strand is called hybridization.

  As shown in FIG. 4A, in the DNA chip 500, a plurality of electrodes 520 are arranged on a substrate 510 made of, for example, glass or silicon, and DNA probes 530 having different arrangements are fixed on each electrode 520. Things. A nucleic acid-amplified specimen sample is caused to flow on the DNA chip 500. At this time, if a DNA probe 530 having a base sequence complementary to the base sequence in the sample sample is present on the DNA chip 500, the two will bind to each other to cause hybridization, thereby generating double-stranded DNA (FIG. 4 (b), (c)). On the other hand, if there is no DNA probe having a base sequence complementary to the base sequence in the sample, no hybridization occurs. Thereafter, the DNA chip 500 is washed, and a reagent (solution) containing the intercalating agent 550 is allowed to flow on the DNA chip 500. Then, the intercalating agent 550 binds to the double-stranded DNA probe 530 where hybridization has occurred. When a voltage is applied to the DNA chip 500 in this state, an oxidizing current of the intercalating agent 550 flows to the electrode 520 to which the double-stranded DNA probe 530 in which hybridization has occurred is immobilized (FIG. 4D).

  FIG. 5A shows an example of this oxidation current, that is, a signal from an electrode where hybridization has occurred, and FIG. 5B shows an example of a signal from an electrode where hybridization does not occur. As can be seen from FIG. 5A, as the voltage applied to the DNA chip 500 increases, the oxidation current sharply increases at a voltage of about 500 mV. On the other hand, the signal value from the electrode where hybridization does not occur slightly increases when the voltage applied to the DNA chip 500 becomes about 500 mV, but the degree of increase is smaller than that in the case shown in FIG. Thus, by determining from which electrode the current has been detected, the sequence of the DNA of the sample can be determined.

  The nucleic acid testing device 110 according to the present embodiment detects the presence or absence of an oxidizing current flowing through the electrode on the DNA chip 500 according to the above-described basic principle, and tests the DNA of the sample.

  In addition, the nucleic acid test of the specimen sample targeted in the present embodiment is not necessarily limited to DNA (deoxyribonucleic acid). The present invention is also applicable to various nucleic acid tests such as RNA (ribonucleic acid), other oligonucleotides and polynucleotides. However, in the following description, an example in which DNA in a specimen sample is tested will be described.

(Nucleic acid test card)
In the nucleic acid testing device 110 according to the present embodiment, a sample is put in the removable nucleic acid testing card 700 and the nucleic acid testing device 110 performs a test.

  FIG. 6 is an external view of the nucleic acid test card 700. The nucleic acid test card 700 is a detachable thin rectangular body, and includes a cap 750, a cover 740, an upper plate 730, a packing 720, a DNA chip 500, and a lower plate 710. Among them, the cap 750, the cover 740, the upper plate 730, and the lower plate 710 are formed of a hard resin member such as PC (polycarbonate), and the packing 720 is formed of an elastic resin member such as an elastomer. Reference numeral 500 is formed of a transparent base material such as glass.

  As described above, since the nucleic acid test card 700 can be composed of only six parts, the material cost and the number of assembly steps can be reduced, and the nucleic acid test card 700 can be provided at low cost. In particular, since the nucleic acid test card 700 according to the present embodiment is premised on being disposable, it is a great advantage that it can be provided at a low price.

  FIG. 7 is a diagram illustrating the procedure for assembling the nucleic acid test card 700. The DNA chip 500 is arranged on the lower plate 710, and the packing 720 is arranged thereon. As shown in FIG. 8A, four recesses 711C1, 711C2, 711C3, and 711C4 are integrally formed on the lower plate 710 in correspondence with the positions of the four syringes. As shown in (b), four dome-shaped protrusions 721C1, 721C2, 721C3, 721C4 are integrally formed in correspondence with the positions of the four syringes. When the packing 720 is mounted on the lower plate 710, as shown in FIG. 8C, the concave portion and the convex portion are arranged to face each other, and four syringes 710C1, 710C2, 710C3, and 710C4 as storage portions are formed. . These syringes 710C1, 710C2, 710C3, and 710C4 individually store sample samples, washing liquids, and the like, as described later.

  As shown in FIGS. 7 and 8A, the lower plate 710 is formed with a groove serving as a flow path for liquid to flow from the syringe 710C1, 710C2, 710C3, 710C4 to the DNA chip 500. . Further, a groove is also formed in the packing 720 in accordance with the groove position of the lower plate 710. Therefore, when the packing 720 is attached to the lower plate 710, a flow path is formed by the grooves arranged to face each other. Further, the upper surface of the DNA chip 500 attached to the lower plate 710 is a flat surface, but a groove is formed at a position of the packing 720 facing the DNA chip 500, and when the packing 720 is attached to the lower plate 710, An inspection channel (inspection section) 712 is formed on the DNA chip 500. The inspection channel 712 is connected to a channel formed by the groove of the lower plate 710 and the groove of the packing 720.

  Next, the upper plate 730 is mounted on the lower plate 710. A part of the upper plate 730 serves as the outer surface of the nucleic acid test card 700. As shown in FIGS. 6 and 7, the upper plate 730 is provided with two through holes 771 and 772 in accordance with the positions of the electrodes of the DNA chip 500. These through holes 771 and 772 are used for inserting the current probe 186 from above and bringing the current probe 186 into contact with the electrodes on the DNA chip 500.

  The upper plate 730 has two through holes 761 and 762 for NO (Normally Open) valves 710a1 and 710a2 for blocking the amplification flow path (amplification unit) 710f of the nucleic acid test card 700 from the flow path connected thereto. Is provided. These through holes 761 and 762 are used for inserting the third rods 241 (third opening and closing unit) and 243 (second opening and closing unit) of the NOV rod 24 to open and close the NO valves 710a1 and 710a2.

  Next, a cover 740 is attached so as to cover a part of the upper plate 730. As shown in FIGS. 6 and 7, the upper plate 730 and the cover 740 are provided with four through holes 781, 782, 783, and 784 in accordance with the positions of the four syringes 710C1, 710C2, 710C3, and 710C4. . These through holes 781, 782, 783, and 784 are for inserting the first rods 201, 202, 203, and 204 of the syringe rod 20 from above to push out the liquid in the syringes 710C1, 710C2, 710C3, and 710C4 to the flow path. Used for

  The upper plate 730 and the cover 740 are provided with four through holes 720d1, 720d2, 720d3, and 720d4 in accordance with the positions of the injection ports of the four syringes 710C1, 710C2, 710C3, and 710C4. The liquid is injected into the four syringes 710C1, 710C2, 710C3, and 710C4 through the through holes 720d1, 720d2, 720d3, and 720d4. More specifically, the specimen sample, the first cleaning solution, the intercalating agent, and the second cleaning solution are respectively supplied to the syringes 710C1, 710C2, 710C3, and 710C4 through separate injection ports 710d1, 710d2, 710d3, and 710d4. Injected. Here, the specimen sample is, for example, a liquid containing any one of an enzyme, deoxyribonucleotide triphosphate (dNTP), a surfactant, magnesium chloride, and ammonium sulfate. The cleaning liquid 1 is a liquid containing, for example, sodium citrate or sodium chloride. The intercalating agent is, for example, a liquid containing Hoechst 33258. The cleaning liquid 2 is a liquid containing, for example, sodium citrate or sodium chloride.

  Further, the upper plate 730 and the cover 740 have four syringes 710C1, 710C2, 710C3, 710C4, and four NC (Normally Close) valves 710v1, which block the flow paths connected to the respective syringes 710C1, 710C2, 710C3, 710C4. Through holes 710h1, 710h2, 710h3, and 710h4 for 710v2, 710v3, and 710V4 are provided. These through holes 710 h 1, 710 h 2, 710 h 3, and 710 h 4 are formed by the second rods 221 (first opening / closing section), 222 (fourth opening / closing section), 223 (fifth opening / closing section), and 224 (sixth opening / closing section) of the NCV rod 23. Is used to open and close the NC valves 710v1, 710v2, 710v3, and 710V4.

  Next, a cap 750 is attached to the cutout of the cover 740 by a hinge, whereby the nucleic acid test card 700 is completed. The reason why the cap 750 is provided is that the user can insert the sample sample from the inlet 710d1 by himself / herself. The first cleaning solution, the second cleaning solution, and the intercalating agent other than the sample sample are previously injected into the corresponding syringe at the stage of manufacturing the nucleic acid test card 700, and the user subsequently injects only the sample sample. Therefore, a cap 750 that covers the injection port 710d1 of the sample sample is provided.

  As described above, the nucleic acid test card 700 according to the present embodiment includes a plurality of syringes 710C1, 710C2, 710C3, and 710C4 for individually storing sample samples and the like, an amplification channel 710f for nucleic acid amplification, and a test flow for DNA test. A path 712 is provided so that nucleic acid amplification and DNA testing can be performed with one card without transferring sample samples, reagents, and the like, so that DNA testing can be automated.

  FIG. 9 is a plan view showing a state in which the cap, the cover 740, and the upper plate 730 have been removed from the nucleic acid test card 700. As shown in FIG. 9, four syringes 710C1, 710C2, 710C3, and 710C4 are arranged on one side of the rectangular nucleic acid test card 700 in the longitudinal direction along the short direction. The syringe (first storage room) 710C1 at the left end stores a sample. The second syringe (second storage room) 710C2 from the left end stores the first cleaning liquid. The second syringe (fourth storage room) 710C3 from the right end stores the intercalating agent. The right end syringe (third storage room) 710C4 stores the second cleaning liquid.

  Beside each syringe 710C1, 710C2, 710C3, 710C4, injection ports 710d1, 710d2, 710d3, 710d4 for injecting liquid into 710C1, 710C2, 710C3, 710C4 in each syringe, and each syringe 710C1, 710C2, 710C. Exhaust holes 710e1, 710e2, 710e3, 710e4 for exhausting the air in 710C4 are provided.

  An amplification channel 710f for amplifying the nucleic acid in the specimen sample is provided at a central portion in the longitudinal direction of the nucleic acid test card 700. The amplification channel 710f and the syringe (710C1) that stores the sample are connected by a channel (first channel). This flow path is branched in the middle, and the branched flow path (third flow path) is connected to a syringe (710C2) for storing the first cleaning liquid. The amplification channel 710f and the inspection channel 712 in the DNA chip 500 are connected by a channel (second channel). Therefore, the sample sample subjected to nucleic acid amplification in the amplification channel 710f can be pushed out by the first washing solution, and the sample sample can be moved to the test channel 712 on the DNA chip 500 without complicated valve control.

  The second flow path is branched in the middle, and a syringe (710C4) that stores the second cleaning liquid is connected to the branched flow path (fourth flow path). Further, this flow path (fourth flow path) is branched in the middle, and a syringe (710C3) for storing the intercalating agent is connected to this branched flow path (fifth flow path). Therefore, the second cleaning liquid stored in the test channel 712 can be pushed out by the intercalating agent.

  NO valves 720a1 and 7120a2 are provided at both ends of the amplification channel 710f. The NO valves 720a1 and 7120a2 are normally open so that the liquid can freely flow between the flow path connected to the amplification flow path 710f and the amplification flow path 710f. When the NO valves 720a1 and 7120a2 are closed, the amplification channel 710f and the channel connected thereto are shut off, and the liquid in the amplification channel 710f flows back to the syringes 710C1, 710C2, 710C3, and 710C4 through the channel. The possibility of flowing into the test channel 712 of the DNA chip 500 is eliminated. In the present embodiment, the NO valves 720a1 and 720a2 are closed while performing nucleic acid amplification in the amplification channel 710f. As a result, the nucleic acid-amplified specimen sample does not mix with the specimen sample in the syringe (710C1).

  FIG. 10 is a schematic sectional view showing the structure of the NO valve. In the packing 720 arranged on the lower plate 710, the location where the NO valve 720a1 is formed is a dome-shaped convex portion, and a flow path is formed inside the convex portion. When this projection is pressed from above by the syringe rod 251, the flow path is closed. The NO valve 720a2 is similarly formed.

  Returning to FIG. 9, NC valves 710v1, 710v2, 710v3, and 710V4 are provided near the connection point between each of the syringes 710C1, 710C2, 710C3, and 710C4 and the flow path. Since there are four syringes 710C1, 710C2, 710C3, and 710C4, four NC valves 710v1, 710v2, 710v3, and 710V4 are also provided. The NC valves 710v1, 710v2, 710v3, and 710V4 are normally closed, and shut off the respective syringes 710C1, 710C2, 710C3, and 710C4 and the flow paths connected thereto. When the NC valves 710v1, 710v2, 710v3, 710V4 are opened, the respective syringes 710C1, 710C2, 710C3, 710C4 are connected to the flow path, and the syringes 710C1, 710C2, 710C3, 710C4 corresponding to the opened NC valves 710v1, 710v2, 710v3, 710V4. The liquid in 710C4 flows into the flow path connected to it.

  11A and 11B are views showing the structure of the NC valve, FIG. 11A is a perspective view, and FIG. 11B is a sectional view taken along line AA of FIG. 11A. The NC valves 710v1, 710v2, 710v3, and 710V4 are cantilever beams formed by a cover 740. The base end of the cantilever is supported by the cover 740, and the tip of the cantilever can move up and down. And the tip is urged in a direction to close the flow path. Therefore, the flow path is normally closed. By pushing up the vicinity of the tip of the cantilever from below by forks 231, 232, 233 and 224 on the tip side of the NCV rod 23, the tip of the cantilever is lifted and the flow path can be opened.

  Returning to FIG. 9, the waste liquid tank 711g1 is arranged along the short side on the side opposite to the side on which the four syringes 710C1, 710C2, 710C3, and 710C4 are arranged with the amplification channel 710f of the nucleic acid test card 700 interposed therebetween. , A DNA chip 500, and a waste liquid tank 711g2. The specimen sample subjected to nucleic acid amplification in the amplification channel 710f flows into the test channel 712 on the DNA chip 500 through the channel. Thereafter, the first cleaning liquid flows into the test channel 712, and the sample in the test channel 712 moves to the waste liquid tank. Next, when the second cleaning liquid flows into the inspection flow path 712, the first cleaning liquid moves to the waste liquid tank, and then, when the intercalant flows into the inspection flow path 712, the second cleaning liquid moves to the waste liquid tank.

  FIG. 12 is a diagram showing a flow channel model of the nucleic acid test card 700. The sample sample, the first cleaning liquid, the intercalating agent, and the second cleaning liquid are respectively injected from the injection ports 710d1, 710d2, 710d3, and 710d4, and stored in the syringes 710C1, 710C2, 710C3, and 710C4. When the NC valve 710v1 is opened, the specimen sample stored in the syringe (710C1) reaches the amplification channel 710f through the channel. Here, the NO valves 720a1 and 7120a2 are closed, and nucleic acid amplification is performed. Thereafter, when the NC valve 710v2 and the NO valves 720a1 and 7120a2 are opened, the first cleaning liquid stored in the syringe (710C2) reaches the amplification channel 710f. The specimen sample that has stayed in the amplification channel 710f until now reaches the test channel 712 on the DNA chip 500 through the channel in such a manner as to be pushed out by the first cleaning solution. After that, when the NC valve 710v4 is opened, the second cleaning liquid stored in the syringe (710C4) reaches the test channel 712 on the DNA chip 500 through the channel. Until now, the sample remaining in the test channel 712 moves to the waste liquid tanks 711g1 and 711g2. Thereafter, when the NC valve 710v3 is opened, the intercalating agent stored in the syringe 710C3 reaches the test channel 712 on the DNA chip 500 through the channel. Until now, the second cleaning liquid remaining in the inspection flow path 712 moves to the waste liquid tanks 711g1 and 711g2.

  FIG. 13 is a plan view showing a detailed structure of the amplification channel 710f. As shown in FIG. 13, the amplification channel 710f is meandering so that the liquid flows slowly. For the nucleic acid amplification, for example, a LAMP (Loop-Mediated Isothermal Amplification) method is used. In the amplification channel 710f, a primer that binds to and amplifies a part of the nucleic acid in the specimen sample is arranged in advance. When a sample is poured into the amplification channel 710f and heated to a predetermined temperature, nucleic acid amplification can be completed in only a few tens of minutes. Amplification products of nucleic acid amplification include not only double-stranded DNA but also single-stranded DNA. In a test channel 712 on the DNA chip 500 described later, current detection is performed using a single strand generated by nucleic acid amplification.

  The technique of nucleic acid amplification according to the present embodiment is not necessarily limited to the LAMP method. Various nucleic acid amplification techniques such as a PCR (Polymerase Chain Reaction) method may be used.

  The amplification flow channel 710f is provided with a plurality of wells at which liquids temporarily accumulate at equal intervals. The present inventor has found for the first time that by providing such a plurality of wells, multi-amplification for amplifying a plurality of different DNAs can be performed. The present inventor has studied and found that the distance between the wells is preferably 4 mm or more along the flow path.

FIG. 14 is a plan view showing the detailed structure of the DNA chip. The DNA chip 500 shown in FIG. 14 includes, on a solid substrate 510, 40 working electrode groups 520 _{1 to} 520 ₄₀ , counter electrodes 522a and 522b, and a reference electrode 524. The counter electrodes 522a and 522b are respectively indicated as CE on the drawing, and the reference electrode 524 is indicated as RE on the drawing. The solid phase substrate 510 is, for example, a glass substrate or a silicon substrate.

The forty working electrode groups 520 _{1 to} 520 ₄₀ are arranged by being divided into first to fourth portions. The first part includes ten working electrode groups 520 _{1 to} 520 ₁₀ and is arranged from left to right on the drawing. The second portion includes ten working electrode groups 520 _{11 to} 520 ₂₀ , which are arranged below the first portion on the drawing and arranged from right to left on the drawing. The third part consists of 10 pairs of working electrodes 520 _{21-520 30} are arranged from left to right in the drawing while being disposed below the second portion in the drawing. The fourth part consists of 10 pairs of working electrodes 520 _{31-520 40} are arranged from right to left in the drawing while being arranged below the third portion in the drawing.

Each working electrode group 520 _i (i = 1,..., 40) has three working electrodes provided at a distance, and a DNA probe having the same DNA sequence is fixed to these working electrodes. Each working electrode is formed of, for example, gold.

Counter electrode 522a has a U-shape, one end of the U-shape are disposed proximate to the working electrode group 520 ₁₀ of the first portion, the other end of the U-shaped operation of the second portion It is arranged close to the electrode group _{520 11.} Counter electrode 522b has a U-shape, one end of the U-shape is disposed proximate to the working electrode group 520 ₃₀ of the third portion, the other end of the U-shaped operation of the fourth portion It is arranged close to the electrode group 520 ₃₁ .

The reference electrode 524 has a U-shape, and one end of the U-shape is arranged close to the working electrode group 520 ₂₀ of the second part, and the other end of the U-shape is operated by the third part. It is arranged close to the electrode groups 520 ₂₁ .

Further, the substrate 510 is provided with terminals W electrically connected to the working electrodes of each working electrode group 520 _i (i = 1,..., 40). A current probe 186 is in contact with these terminals W.

The substrate 510 further includes C terminals 523a and 523b, R terminals 525a and 525b, an X terminal 526, Y terminals 528a and 528b, and a plurality of conduction detection terminals 58. C terminal 523a is electrically connected to counter electrode 522a, and C terminal 523b is electrically connected to counter electrode 522b. The R terminals 525a and 525b are connected to the reference electrode 524, respectively. The X terminals 526a and 526b are provided at both ends of the DNA chip 500, and are terminals for applying a voltage between these terminals 526a and 526b to detect whether the DNA chip 500 is conductive. Similarly, Y terminals 528a and 528b are provided at both ends of the DNA chip 500, and are terminals for applying a voltage between these terminals 528a and 528b and detecting whether or not the DNA chip 500 is conductive. The plurality of conduction detection terminals 580 are provided corresponding to the working electrodes of the working electrode groups 520 _{1 to} 520 ₄₀ , and each conduction detection terminal 580 is connected to the corresponding working electrode and electrically connected to the corresponding working electrode. By applying a voltage between the terminal W and the terminal W, it is detected whether or not the connection between the corresponding working electrode and the terminal W is conducted. As shown in FIG. 6, the terminals are provided in upper and lower sets on the drawing so as to sandwich the working electrode group 520 _i (i = 1,..., 40).

The DNA chip 500 is mounted on the nucleic acid test card 700. When mounted, to flow reagents containing intercalating agent toward the working electrode 520 ₁₀ from the working electrode group 520 _1, the flow path is formed on the first portion. Further, also on the counter electrode 522a, the flow path to flow is the reagent toward the working electrode _{520 11} from the working electrode group _{520 10} is formed. As flows the reagent toward the working electrode 520 ₂₀ from the working electrode group 520 _11, the flow path is formed on the second portion. Also, a flow path is formed on the reference electrode 524 so that the reagent flows from the working electrode group 520 ₂₀ toward the working electrode 520 ₂₁ . A flow path is formed on the third portion so that the reagent flows from the working electrode group 520 ₂₁ toward the working electrode 520 ₃₀ . Further, also on the counter electrode 522b, the flow path to flow is the reagent toward the working electrode _{520 31} from the working electrode group _{520 30} is formed. A flow path is formed on the fourth portion so that the reagent flows from the working electrode group 520 ₃₁ toward the working electrode 520 ₄₀ . These channels are formed by the DNA chip 500 and the packing 720 of the nucleic acid test card 700.

  The reagent containing the intercalating agent includes a channel on the first portion, a channel on the counter electrode 522a, a channel on the second portion, a channel on the reference electrode 524, a channel on the third portion, It passes through the flow path on the counter electrode 522b and the flow path on the fourth portion in this order.

(Configuration and operation of nucleic acid testing device)
FIG. 15 is a block diagram of a control system of the nucleic acid testing apparatus 110 according to the present embodiment. As shown in FIG. 15, the control system of the nucleic acid testing apparatus 110 includes a control unit 151, a communication unit 152, a tray presence / absence sensor 153, a card presence / absence sensor 154, a housing fan 155, a buzzer 156, and an LED 157. , A current probe control unit 158, a motor group 159, a photo sensor 160, a temperature sensor 161, a temperature adjustment unit 162, and a power supply unit 163.

  The communication unit 152 performs data communication between the information processing device 150 and the control unit 151. For example, the communication unit 152 receives information set on the GUI screens 201, 202, 203, and 204 displayed on the screen of the information processing device 150 from the information processing device 150 and transmits the information to the control unit 151. In addition, the communication unit 152 transmits the detection result of the current flowing to the electrode of the DNA chip 500 from the control unit 151 to the information processing device 150, and transmits the abnormality detected by the control unit 151 from the control unit 151 to the information processing device 150. Transmit. As a communication interface of the communication unit 152, for example, USB can be used, but a specific communication interface does not matter. In addition, the communication unit 152 does not necessarily need to be wired, and may perform wireless data communication.

  The tray presence / absence sensor 153 detects opening / closing of the trays 111, 112, 113, 114 and transmits the detection result to the control unit 151. The tray presence / absence sensor 153 detects opening and closing of the trays 111, 112, 113, 114, for example, mechanically or optically.

  The card presence / absence sensor 154 detects whether or not the nucleic acid test card 700 has been placed on the trays 111, 112, 113, 114, and transmits the detection result to the control unit 151. The detection of whether or not the nucleic acid test card 700 is placed on the trays 111, 112, 113, 114 can be performed by, for example, a sensor that mechanically or optically detects the presence or absence of the nucleic acid test card 700.

  As shown in FIG. 16, for example, two housing fans 155 are provided along the vertical direction on the rear side of the housing placed vertically (up and down) from the installation surface. An air inlet is provided on the lower front side of the housing. The rotation / stop of the housing fan 155 is controlled by the control unit 151. When the control unit 151 rotates the housing fan 155, outside air is taken into the housing from the air inlet, circulated in the housing, and then exhausted from the housing fan 155. Thereby, the entire inside of the housing is cooled. The number and location of the housing fans 155 may be arbitrarily changed according to the size and shape of the housing.

  The buzzer 156 and the LED 157 are turned on, for example, when any abnormality occurs in the nucleic acid testing device 110. The sounding state of the buzzer 156 and the lighting mode of the LED 157 may be switched depending on the type of abnormality. The control unit 151 turns on / off the buzzer 156 and the LED 157.

  The current probe control unit 158 detects a current flowing through the plurality of current probes 186 that are in contact with the plurality of electrodes of the DNA chip 500. Since the current flowing through each current probe 186 is small, the current probe control unit 158 performs a process of removing the noise, amplifying the current flowing through each current probe 186, and converting the current into a voltage.

  The motor group 159 has five motors 181, 182, 183, 184, and 185. Among these motors, the syringe shaft motor 184 controls the driving of the syringe rods 201, 202, 203, and 204 that move the liquid in each syringe to the flow path. The NOV shaft motor 181 controls the driving of the NOV rod 24 that opens and closes the NO valves 710a1 and 710a2. The NCV shaft motor 185 controls the driving of the NC rod 20 that opens and closes the NC valves 710v1, 710v2, 710v3, and 710V4. The heater / Peltier shaft motor 183 controls the driving of the temperature control support 25 on which the heater 170 and the Peltier element 171 are mounted. The probe shaft motor 182 controls driving of the probe support 26 to which the current probe 186 is attached.

  Photosensors 160 are arranged near the five motors 181, 182, 183, 184, and 185, respectively. The photo sensor 160 corresponding to each motor optically detects the operation origin position of each motor and performs an operation of returning each motor to the operation origin position.

  As shown in FIG. 17, the temperature adjustment unit 162 separates the temperatures of the heater 170 and the Peltier element 171 based on the temperature detection results of the temperature sensors 161a and 161b provided near the heater 170 and the Peltier element 171, respectively. Adjust to In addition, a temperature sensor 161c for measuring the temperature inside the housing is provided separately from the temperature sensors 161a and 161b. The temperature adjustment unit 162 controls the strength of the housing fan 155 based on the temperature inside the housing. On / off is instructed to the control unit 151.

  The power supply unit 163 generates a plurality of DC power supply voltages used in each unit in the nucleic acid test apparatus 110 and supplies the DC power supply voltages to the units. The power supply unit 163 has an AC / DC converter that generates a plurality of power supply voltages from a commercial power supply.

  FIG. 18 shows the arrangement of the five motors 181, 182, 183, 184 and 185 described above and the syringe rod 20, NCV rod 23, NOV rod 24, temperature control support 25, and probe support 26 driven by these motors. FIG. FIG. 18 is a plan view seen from the side of the nucleic acid testing apparatus 110, where the right side in the figure is the front and the left side is the back. As shown in FIG. 18, a NOV axis motor 181, a probe axis motor 182, and a heater / Peltier axis motor 183 are supported on a support plate 180 extending in a vertical (vertical) direction in order from top to bottom. A support plate 191 for supporting the syringe shaft motor 184 is disposed vertically above the front side. Further, below the front side, a support plate 192 that supports the NCV axis motor 185 is disposed in the vertical direction. The rotating shafts of these five motors are all arranged in the horizontal direction. A rack gear 14d as shown in FIG. 19 meshes with the gears 14c of the rotating shafts, and the rotation of the rotating shaft 12c is converted into a linear motion in the vertical direction, and the rod corresponding to each motor moves in the vertical direction. . Further, by switching the rotation direction of the rotation shaft of each motor, the corresponding rod moves upward or downward.

  FIG. 20 is a plan view showing the syringe rod 20 driven by the syringe shaft motor 184. The syringe rod 20 has four first rods 201, 202, 203, and 204 that come into contact with the four syringes 710C1, 710C2, 710C3, and 710C4 in the nucleic acid test card 700. These four first rods 201, 202, 203, and 204 extend downward from above the housing, and the heights of the distal ends of the first rods are different from each other. More specifically, the left end of the first rod 201 is located at the lowest position, the second end of the first rod 202 from the left end is low, and the end position of the right end first rod 204 is next. And the tip position of the second rod 203 from the right end is the highest. These four first rods 201, 202, 203, and 204 are supported by a common support plate 205, and the support plate 205 moves up and down with the rotation of the rotation shaft of the syringe shaft motor 184. Therefore, the four first rods 201, 202, 203, 204 move up and down in synchronization.

  Resilient punches 211, 212, 213, 214 are attached to the tips of the four first rods 201, 202, 203, 204.

  FIG. 21 is a perspective view of the punch 213. The punch 213 has an elongated shape in accordance with the shape of the syringe, and presses the upper surface of the dome-shaped packing 720 constituting the syringes 710C1, 710C2, 710C3, and 710C4 from above to push the liquid in the syringe to the corresponding flow path. .

  In order to specify the shape of the punch, the present inventor prepared two types of punches having different widths, and examined whether or not liquid remains in the syringe when the syringe was pressed by the punch. FIG. 22A shows an example in which a wide punch 213a is pressed against a syringe 710c3, and FIG. 22B shows an example in which a narrow punch 213b is pressed against a syringe 710c3. In addition, FIG. 23 shows a case where the liquid amount of the syringe 710c3 when the position where the punch 213b presses the syringe 710c3 is shifted is shown in FIG. 22A, and the packing 720 of the syringe 710c3 is pressed when the punch 213a is pressed against the syringe 710c3. As shown in the figure, the packing deformed into an M shape, a large shear load occurred between the packings, and liquid remained in the syringe. On the other hand, in the case of FIG. 22B, since the contact area between the punch 213b and the syringe 710c3 is small, stress is concentrated at the center of the syringe 710c3, and it is understood that no liquid remains in the syringe 710c3. Was. More specifically, it was found that it is desirable that the width in the short direction of the punch be smaller than the width in the short direction of the syringe 710c3 by 0.3 mm or more.

  Further, as shown in FIG. 23, when the syringe 710C is pushed by the syringe rod 251 at a position deviated from the center of the syringe 710C, the syringe 710C cannot be sufficiently pushed in, so that a sufficient liquid sending amount cannot be secured. Therefore, as shown in FIG. 24, a portion that does not touch the nucleic acid test card 500 above the tip of the syringe rod 251 that moves up and down along the rod guide 252 is formed into a tapered structure 253, and the position is aligned with the nucleic acid test card 700. May be made. With this structure, it is possible to avoid a decrease in the amount of liquid transfer due to a shift in the position where the syringe rod 251 presses the syringe 710C, and thus an appropriate amount of liquid transfer can be realized.

  The four first rods 201, 202, 203, and 204 are formed of a resilient member such as a spring, and have punches 211, 212, 213, and 214 attached to tips thereof. In the normal state, each of the first rods has an urging force acting downward, and as shown in FIG. 20, the heights of the tip portions are different from each other. When the support plate is lowered, the punch 211 of the first rod whose tip is at the lowest position is pressed against the upper surface of the corresponding syringe (710C1) first, and the first rod 201 contracts. The lower the support plate 205 is, the more the punch 211 is pressed against the syringe 710C1, and all the liquid inside the syringe 710C1 is pushed out to the corresponding flow path.

  Since the heights of the tip portions of the four first rods 201, 202, 203, and 204 are different from each other, first, the first rod 201 on the left end contacts the syringe 710C1, and then the second first rod 202 from the left. Comes into contact with the syringe 710C2, then the first rod 204 at the right end contacts the syringe 710C4, and finally, the first rod 203 from the right end contacts the syringe 710C3. The liquid in the syringe is extruded from the syringe to the corresponding flow path in the order in which the punch contacts the upper surface of the syringe.

  As described above, in the nucleic acid test card 700 of the present embodiment, it is necessary to move the syringe 710C1, 710C2, 710C3, 710C4 to the corresponding flow path in the order of the sample sample, the first cleaning liquid, the second cleaning liquid, and the intercalating agent. There is. Therefore, in FIG. 20, the heights of the tip portions of the four first rods are made different according to the order in which the liquid is sent from the syringes 710C1, 710C2, 710C3, and 710C4 to the corresponding flow paths. The liquid in the syringes 710C1, 710C2, 710C3, and 710C4 can be moved to the respective flow paths in a predetermined order simply by moving the liquid once from the upper side to the lower side, and the drive control of the syringe rod is facilitated.

  FIG. 25 is a plan view showing the NCV rod 23 driven by the NCV shaft motor 184. The NCV rod 23 has four second rods 221, 222, 223, and 224 that open and close four NC valves 710v1, 710v2, 710v3, and 710V4 in the nucleic acid test card 700. These four second rods extend upward from the bottom of the housing, and fork portions 231, 232, 233, which are forked at the distal ends of the second rods 221, 222, 223, 224. 234 are provided. The heights of the forks 231, 232, 233, 234 of these four second rods are different from each other, and each of the second rods 221, 222, 223, 224 has a left end 221, a second end 222 from the left end, and a right end 224. , The height becomes lower in the order of the second 223 from the right end.

  The tip of the fork portion 231, 232, 233, 234 of the second rod has a curved surface shape (R surface shape) as shown in FIG. 26 (a) or FIG. 26 (b), or as shown in FIG. 26 (c). It is preferable to have a tapered shape (C-plane shape). With such a tip shape, each NC valve can be opened and closed even if the position of each NC valve in the nucleic acid test card 700 is slightly shifted.

  Each of the NC valves 710v1, 710v2, 710v3, and 710V4 has a cantilever structure as described with reference to FIG. 11, and the tip of the cantilever exerts an urging force in a direction to close the flow path. The fork portions 231, 232, 233, 234 on the tip side of the second rod lift the tip portion of the cantilever upward from both sides to open the flow path.

  The four second rods 221, 222, 223, and 224 are supported by a common support plate 230, and the support plate 230 moves up and down with the rotation of the rotation shaft of the NCV shaft motor 184. Therefore, the four second rods 221, 222, 223, and 224 move up and down in synchronization. Since the second rods 221, 222, 223, and 224 move upward from below, the fork portions 231, 232, 233, and 234 of the second rods are connected to the corresponding NC valves 710v1, 710v2, 710v3, and 710V4. The holding valve is lifted upward by contacting the holding beam from below, and the NC valves 710v1, 710v2, 710v3, and 710V4 are opened. When the second rod moves downward from above and stops contacting the NC valves 710v1, 710v2, 710v3, and 710V4, the NC valve returns to the original closed state.

  The four second rods 221, 222, 223, 224 are formed of a resilient member such as a spring, and when the tips contact the NC valves 710 v 1, 710 v 2, 710 v 3, 710 V 4, the second rods 221. , 222, 223, and 224 contract, and when their distal ends are separated from the NC valves 710v1, 710v2, 710v3, and 710V4, they return to the initial position shown in FIG.

  When the NCV axis motor 184 is rotated to move the support plate 230 upward, first, the second rod 221 on the left end opens the corresponding NC valve 710v1, and the sample flows into the amplification channel 710f. When the support plate 230 is further raised, the second rod 222 from the left end opens the corresponding NC valve 710v2, and the first cleaning liquid flows into the amplification channel 710f. When the support plate 230 is further raised, the second rod 224 on the right end opens the corresponding NC valve 710V4, and the second cleaning liquid flows through the flow path into the test flow path 712 on the DNA chip 500. When the support plate 230 is further raised, the second rod 223 from the right end opens the corresponding NC valve 710v3, and the intercalant flows into the test channel 712 on the DNA chip 500 through the channel.

  As described above, the four second rods 221, 222, 223, and 224 are supported by the common support plate 230 and moved up and down in a state where the heights of the tip portions are different from each other. By moving upward, the four NC valves 710v1, 710v2, 710v3, and 710V4 can be sequentially opened, and the liquid in the four syringes 710C1, 710C2, 710C3, and 710C4 can be sequentially moved to the corresponding flow paths. Therefore, the opening / closing control of the NC valves 710v1, 710v2, 710v3, and 710V4 can be easily performed.

  FIG. 27 is a plan view showing the NOV rod 24 driven by the NOV shaft motor 181. The NOV rod 24 has two third rods 241 and 243 for opening and closing the two NO valves 710a1 and 710a2 in the nucleic acid test card 700, and a third rod 242 for positioning. These three third rods 241, 242, 243 extend downward from above the housing. Of the three, the third positioning rod 242 at the center is larger than the remaining two rods 241, 243. The height of the tip is slightly higher.

  The third rods 241, 242, 243 are supported by a common support plate 240, and the support plate 240 moves up and down with the rotation of the rotation shaft of the NOV shaft motor 181. Therefore, the three third rods 241, 242, 243 move up and down in synchronization.

  When the support plate 240 is lowered, the center third rod 242 among the three first contacts the upper surface of the nucleic acid test card 700 to perform positioning. As shown in FIG. 6, the remaining two third rods 241 and 243 press the tube structure of the packing 720 from above to close the flow path.

  As shown in FIG. 27, the positioning third rod 242 has a tip position higher than the remaining two third rods 241 and 243, but these two third rods 241 and 243 are Since the gasket 720 is in contact with the packing 720 below the upper surface of the nucleic acid test card 700, the third positioning rod 242 contacts the upper surface of the nucleic acid test card 700 before the two third rods 241 and 243. Then, positioning is performed.

  As described in FIG. 9, the two NO valves 710a1 and 710a2 are provided at the entrance and exit of the amplification flow path 710f on the nucleic acid test card 700, and these NO valves 710a1 and 710a2 are connected to the third rods 241 and 243. At the same time, it is possible to prevent the liquid in the amplification channel 710f from flowing back to the syringe or flowing to the DNA chip 500 side. In the present embodiment, nucleic acid amplification is performed by heating the amplification channel 710f with the NO valves 710a1 and 710a2 closed.

  FIG. 28A is a plan view showing the temperature control support 25 driven by the heater / Peltier shaft motor 183, and FIG. 28B is a perspective view of the temperature control support 25. The temperature control support 25 includes a first support plate 174 that supports the heater 170, a second support plate 175 that supports the Peltier element 171, a first support plate 174, a second support plate 175, and a third support plate 176. And a telescopic member such as a spring disposed between them. The first support plate 174 and the second support plate 175 are arranged at substantially the same height when the elastic member is not urged. The third support plate 176 moves up and down by the rotation 183 of the heater / Peltier shaft motor, and accordingly, the first support plate 174 and the second support plate 175 also move up and down in synchronization.

  The heater 170 disposed on the upper surface of the first support plate 174 has a rectangular shape, and the Peltier element 171 disposed on the upper surface of the second support plate 175 has a rectangular shape. When the third support plate 176 moves upward, the heater 170 is housed in one recess provided on the back surface side of the nucleic acid test card 700, and the Peltier element 171 is housed in the other recess. The concave portion on the back surface side of the nucleic acid test card 700 has a size corresponding to the outer shape of the heater 170 and the Peltier device 171, and the heater 170 and the Peltier device 171 are housed in these concave portions, so that the heater 170 and the Peltier device 171 are formed. Accurate positioning is performed. The concave portion for the heater in the nucleic acid test card 700 is provided directly below the amplification channel 710f, and the concave portion for the Peltier element is provided directly below the test channel 712 on the DNA chip 500. Thus, the heater 170 can heat the amplification channel 710f, and the Peltier element 171 can heat and cool the inspection channel 712.

  As described above, the heater 170 and the Peltier device 171 come into contact with the nucleic acid test card 700 almost simultaneously, but the heating control of the heater 170 and the temperature control of the Peltier device 171 can be performed individually. In the present embodiment, before starting nucleic acid amplification with the nucleic acid test card 700, the temperature control support 25 is moved upward to bring the heater 170 and the Peltier element 171 into contact with the nucleic acid test card 700, and thereafter, the inside of the syringe The sample sample is moved to the amplification channel 710f, and nucleic acid amplification is performed while heating with a heater. Thereafter, when the nucleic acid-amplified specimen sample moves to the test channel 712 on the DNA chip 500, the test channel 712 is heated and cooled by the Peltier device. The temperature control support 25 is kept moving upward until the probe is brought into contact with the DNA chip 500 and current detection is performed.

  FIG. 29A is a plan view showing the probe support 26 driven by the probe shaft motor 182, and FIG. 29B is a perspective view of the probe support 26. The probe support 26 is arranged above the nucleic acid test card 700. The probe support 26 has a support plate 197 that supports the current probe 186. The support plate 197 moves up and down according to the rotation of the rotation shaft of the probe shaft motor 182. On the back side of the support plate 197, a current probe 186 and pins 261 and 262 for positioning the current probe 186 are provided. The current probe 186 has a structure in which a spring 263 is built in, as shown in FIG. When the pressing pressure of the pins 261 and 262 is small, there is a possibility that conduction cannot be obtained due to the contact resistance. However, by providing the spring 263, the pressing pressure of the probe pins 261 and 262 is reduced to the conduction with respect to the nucleic acid test card 700. Possible indentation stresses can be achieved. In the present embodiment, the spring 263 is built in, but an elastic body other than the spring 263 can be appropriately used.

  The current probes 186 are provided by the number of electrodes on the DNA chip 500, and a wiring pattern for detecting a current flowing through each current probe 186 is formed on the support plate 197. When the probe shaft motor 182 is driven, the current probe 186 gradually descends from above, and first, the positioning pins 261 and 262 are fitted into the holes on the nucleic acid test card 700 to be positioned. The probe 186 contacts each electrode on the DNA chip 500. Since the positioning is performed first by the positioning pins 261 and 262, the large number of current probes 186 accurately contact the electrodes on the corresponding DNA chip 500.

  FIG. 31 is a flowchart illustrating an example of a drive sequence of each motor by the control unit 151, and FIG. 32 is a diagram illustrating a moving state of a liquid on the nucleic acid test card 700. Hereinafter, the procedure of the nucleic acid test will be described with reference to these drawings.

  The probe shaft motor 182 is driven to lower the probe support 26. Then, with the current probe 186 positioned, the current probe 186 is brought into contact with each electrode on the DNA chip 500 mounted on the nucleic acid test card 700 (step S1).

  The NOV axis motor 181 is driven to bring the third rod 242 for positioning the NOV rod 24 into contact with a predetermined position on the nucleic acid test card 700 to perform positioning (step S2).

  The heater / Peltier shaft motor 183 is driven to move the temperature control support 25 upward, and the heater 170 and the Peltier element 171 are both housed in the concave portion on the back side of the nucleic acid test card 700 for positioning (step S3). . In this state, since the NC valves 710v1, 710v2, 710v3, and 710V4 are all closed, the liquid in the four syringes 710C1, 710C2, 710C3, and 710C4 flows into the flow path as shown in FIG. Absent.

  The NC valve shaft motor 185 is driven to raise the NCV rod 23, and at the tip of the second rod 221 whose tip is at the highest position, the corresponding cantilever is lifted, and the syringe for the sample sample is lifted. The NC valve 710v1 at the outlet of is opened (step S4).

  By driving the syringe shaft motor 187, the syringe rod 20 is lowered, and the first rod 201 whose tip is at the lowest position is pressed against the upper surface of the sample sample syringe 710C1 to push the sample sample into the flow path. (Step S5). As a result, as shown in FIG. 32B, the sample in the sample sample syringe 710C1 flows through the flow channel to the amplification flow channel 710f.

  The NOV shaft motor 181 is driven to bring the two third rods 241 and 243 of the NOV rod 24 into contact with the packing 720 of the NO valve to close the NO valves 710a1 and 710a2 and to use the heater 170 to amplify the flow path 710f. Is heated (step S6). By this heating, the nucleic acid contained in the specimen sample is amplified in the amplification channel 710f. When the nucleic acid amplification is completed, the heating by the heater 170 is stopped. Since the heater 170 does not particularly have a cooling function, it is naturally cooled.

  The NOV shaft motor 181 is driven in the direction opposite to the step S6, and the NO valves 710a1 and 710a2 are opened (step S7).

  The NC valve shaft motor 185 is driven in the same direction as that in step S4 to further raise the NCV rod 23, and the corresponding cantilever is provided at the tip of the second rod 222 whose tip is at the second highest position. Is lifted, and the NC valve 710v2 at the outlet of the syringe 710C2 for the first cleaning liquid is opened (Step S8).

  The syringe shaft motor 187 is driven in the same direction as in step S5 to further lower the syringe rod 20 and press the first rod 202 whose tip is at the second lowest position against the upper surface of the syringe 710C2 for the first cleaning liquid. Then, the first cleaning solution is pushed out to the amplification channel 710f (Step S9). Thereby, as shown in FIG. 32 (c), the first washing liquid enters the amplification channel 710f, and the sample sample subjected to nucleic acid amplification in the amplification channel 710f is pushed out to the detection channel in the DNA chip 500.

  Similarly to step S6, the NOV shaft motor 181 is driven to bring the two third rods 241 and 243 of the NOV rod 24 into contact with the packing 720 of the NO valve again, and the NO valves 710a1 and 710a2 are closed. In parallel with this, the DNA chip 500 is heated by the Peltier device 171 (step S10).

  Next, the NC valve shaft motor 185 is driven in the same direction as in steps S4 and S8 to further raise the NCV rod 23, and at the distal end of the second rod 224 whose distal end is at the third highest position, The corresponding cantilever is lifted, and the NC valve 710v4 at the outlet of the second cleaning liquid syringe 710C4 is opened (step S11).

  Next, the syringe shaft motor 184 is driven in the same direction as in steps S5 and S9 to further lower the syringe rod 20, and the first rod 204 whose tip is located at the third lowest position is used as the syringe for the second cleaning liquid. The second cleaning liquid is pushed out onto the flow path connected to the DNA chip 500 by pressing it against the upper surface of the 710C4 (step S12). As a result, as shown in FIG. 32 (d), the second cleaning liquid moves to the test flow path 712 on the DNA chip 500, and the sample stored in the test flow path 712 up to that time is transferred to the waste liquid tanks 711g1 and 711g2. Extruded.

  Next, the NC valve shaft motor 185 is driven in the same direction as in steps S4, S8, and S11 to further raise the second rod, and the tip of the second rod 223 whose tip is at the lowest position is moved. Then, the corresponding cantilever is lifted, and the NC valve 710v3 at the outlet of the syringe 710C3 for the insertion agent is opened (step S13).

  Next, the syringe shaft motor 187 is driven in the same direction as in steps S5, S9, and S12 to further lower the syringe rod 20, and the first rod 203 having the highest distal end is connected to the syringe for the insert. The insertion agent is pressed against the upper surface of the 710C3 to push out the intercalating agent into the flow path connected to the DNA chip 500 (step S14). As a result, as shown in FIG. 32 (e), the intercalating agent moves to the test channel 712 on the DNA chip 500, and the second cleaning liquid that has accumulated in the test channel 712 is pushed out to the waste liquid tank. Thereafter, a current flowing through the electrode is detected by a current probe 186 in contact with the electrode of the DNA chip 500.

  As described above, in the nucleic acid testing apparatus 110 according to the present embodiment, the user simply places the nucleic acid testing card 700, which can perform both nucleic acid amplification and nucleic acid testing, on the tray of the nucleic acid testing apparatus 110, and then automatically performs nucleic acid amplification and nucleic acid testing. Therefore, not only can the labor of the nucleic acid test be omitted and the convenience can be improved, but also the time until the nucleic acid test result is obtained can be greatly reduced.

  In addition, a single nucleic acid test card 700 can store all sample samples and necessary reagents and the like, and since both nucleic acid amplification and nucleic acid tests can be performed in this card, the amount of reagents and the like to be used can be reduced. The required member costs can be greatly reduced.

  Note that, in order to perform positioning with higher accuracy, the following step may be added as a step before S1 “unload and position the current probe 186” in FIG.

  (1) First, in order to roughly position the nucleic acid test card 700 and the probe support 26, the probe shaft motor 182 is driven to lower the probe support 26, and as shown in FIG. Are inserted into the positioning holes of the nucleic acid test card 700, and the probe support 26 is stopped before the current probe 186 contacts the corresponding electrode.

  (2) Next, in order to further perform positioning, the NC valve rod motor 185 is driven to raise the NCV rod 23, and the tip of the second rod 221 corresponding to the highest position corresponds to the tip. Stop NC valve shaft motor 185 before contacting the cantilever.

  By adding the above pre-process, more accurate positioning can be realized.

  As shown in FIG. 18, the nucleic acid testing device 110 has a compact housing of necessary motors and the like in a small space housing. Further, as shown in FIG. And power consumption can be suppressed.

  At least a part of the nucleic acid testing apparatus 110 described in the above embodiment may be configured by hardware or software. When configured with software, a program for realizing at least a part of the function of the nucleic acid testing device 110 may be stored in a recording medium such as a flexible disk or a CD-ROM, and may be read and executed by a computer. The recording medium is not limited to a removable medium such as a magnetic disk or an optical disk, but may be a fixed recording medium such as a hard disk device or a memory.

  Further, a program that realizes at least a part of the function of the nucleic acid testing device 110 may be distributed via a communication line (including wireless communication) such as the Internet. Further, the program may be encrypted, modulated, or compressed, and distributed via a wired or wireless line such as the Internet, or stored in a recording medium.

  Although several embodiments of the present invention have been described, these embodiments are provided by way of example and are not intended to limit the scope of the invention. These new embodiments can be implemented in other various forms, and various omissions, replacements, and changes can be made without departing from the spirit of the invention. These embodiments and their modifications are included in the scope and gist of the invention, and are also included in the invention described in the claims and their equivalents.

110: nucleic acid testing device, 111, 112, 113, 114: tray, 150: information processing device, 170: heater, 171: Peltier element, 181, 182, 183, 184, 185 ... Motor, 186 ... Current probe, 201, 202, 203, 204 ... 1st rod, 221, 222, 223, 224 ... 2nd rod, 241, 242, 243 ... 3rd Rod, 500: DNA chip, 520: Electrode (working electrode), 700: Nucleic acid test card, 710a1, 710a2: NO valve, 710C1, 710C2, 710C3, 710C4: Syringe, 710f ..Amplification channels, 710v1, 710v2, 710v3, 710V4... NC valves, 712...

Claims (6)

A storage unit that stores at least a sample sample, an amplification unit that amplifies nucleic acids contained in the sample sample stored in the storage unit, a first flow path that moves the sample sample from the storage unit to the amplification unit, A testing unit for testing nucleic acids contained in the sample sample amplified by the amplifying unit, a second flow path for moving the sample sample from the amplifying unit to the testing unit, and the first washing liquid from the storage unit to the amplifying unit. And a third flow path branched from the first flow path; and a mounting section for mounting a nucleic acid testing device,
A first opening / closing unit that is provided in the first flow path and controls outflow of a sample from the storage unit;
A second opening / closing unit that is provided in the second flow path and controls the flow of the sample sample into the test unit;
A third opening / closing unit that is provided in the first flow path and controls the flow of the sample to the amplification unit;
A fourth opening / closing unit that opens and closes the third flow path;
A heating unit for heating the amplification unit;
A heating and cooling unit for adjusting the temperature of the inspection unit,
After opening the first opening / closing section and moving the sample sample stored in the storage section to the amplification section, the second opening / closing section and the third opening / closing section are closed, and the amplifying section is heated by the heating section. Is heated to perform nucleic acid amplification in the amplification section, and after the nucleic acid amplification in the amplification section is completed, both the second to fourth opening / closing sections are opened, and the first washing liquid is supplied to the third flow path and the third flow path. Along with moving to the amplifying section via the first flow path, the sample sample subjected to nucleic acid amplification in the amplifying section is moved to the test section via the second flow path, closing the second opening / closing section, A control unit that performs temperature adjustment of the inspection unit in a heating and cooling unit , and the amplification unit includes a plurality of wells that amplify a plurality of types of nucleic acids.
Nucleic acid testing device. The storage unit further stores a second cleaning solution,
The nucleic acid testing device further includes a fourth flow path branched from the second flow path, which moves the second cleaning liquid from the storage unit to the testing unit,
A fifth opening / closing unit that opens / closes the fourth flow path;
Wherein the control unit, the fifth opening the opening and closing unit, nucleic acid testing apparatus according to claim 1 for moving the second cleaning liquid to the measurement part via the fourth flow path and the second flow path. The storage unit further stores a reagent used for nucleic acid testing,
The nucleic acid testing device further comprises: a fifth channel separated from the second channel, which moves the reagent from the storage unit to the testing unit,
A sixth opening / closing unit that opens / closes the fifth flow path,
The control unit opens the sixth opening / closing unit to move the reagent to the test unit via the fifth flow path and the second flow path, and the nucleic acid contained in the sample sample in the test unit The nucleic acid testing device according to claim 2 , wherein the testing is performed. The nucleic acid testing device has a plurality of electrodes to each of which a plurality of probe molecules that cause hybridization between different types of nucleic acids are attached,
4. The test section according to claim 3 , wherein the test section tests nucleic acids contained in the specimen sample by checking whether or not an oxidation current due to hybridization flows through the plurality of electrodes when the reagent is moved to the test section. Nucleic acid testing device. The storage unit includes a first storage chamber for storing the sample sample, a second storage chamber for storing the first cleaning liquid, a third storage chamber for storing the second cleaning liquid, and a fourth storage chamber for storing the reagent. And a storage room,
Each of the first to fourth storage chambers is provided with four first rods for applying pressure to the storage chamber and extruding the storage material into a corresponding flow path,
The control unit synchronously moves these four first rods in one direction,
While the four first rods are moving in the one direction, one of the four first rods first applies pressure to the first storage chamber to cause the sample to flow through the first storage chamber. Path, and then another first rod applies pressure to the second storage chamber to move the first cleaning solution to the third flow path, and then another first rod is moved to the third storage chamber. Applying pressure to the second cleaning liquid to move to the fourth flow path, and then another first rod applying pressure to the fourth storage chamber to move the reagent to the fifth flow path. 5. The nucleic acid testing device according to 3 or 4 . The second and third opening / closing portions open / close a first valve which is in an open state in a normal case where no pressure is applied and which is closed when a pressure is applied,
Said first and fourth to sixth switching unit, if the normally not apply pressure in the closed state, according to claim 3 or 4 for opening and closing the second valve to be opened when a pressure is applied Nucleic acid testing device.

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