JP2011214907A - Energization test cassette used for energization test of nucleic acid concentration quantitative analyzing apparatus, and energization test method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To accurately conduct an energization test of a current detection type nucleic acid concentration quantitative analyzing apparatus in an inexpensive manner in a short time.SOLUTION: An analyzing apparatus 60 includes a probe pin 82, a detection means 90, and an analysis means 100. The analyzing apparatus 60 is loaded with an analysis cassette incorporating a DNA chip and obtains a concentration of a specimen DNA immobilized on the DNA chip. The energization test of the detection means 90 is conducted by loading the analyzing apparatus 60 with an energization test cassette. The energization test cassette contains a plurality of current detection pads, a plurality of resistance parts connected to the current detection pads and having mutually different resistance values, and a resistance circuit board on which voltage application pads connected to the resistance parts are formed. The energization test of the detection means 90 is conducted by abutting the probe pin 82 against the voltage application pad, applying a voltage to the resistance part to detect a current value from the current detection pad, and comparing a resistance value calculated based on the detected current value and an actual resistance value of the resistance part.

Description

  The present invention relates to a current-carrying test cassette used for a current-carrying test of a nucleic acid concentration-quantitative analyzer provided in a current detection type nucleic acid concentration-quantitative analysis system, and a current-carrying test method using the same.

  The current detection type nucleic acid concentration quantitative analysis system includes a nucleic acid concentration quantitative analysis cassette in which a DNA chip is incorporated, and a nucleic acid concentration quantitative analysis device in which the analysis cassette 30 is mounted.

  The DNA chip is formed with a plurality of working electrodes, a plurality of counter electrodes to which a voltage is applied between each working electrode, and a plurality of working electrodes and a plurality of measuring electrodes connected to each counter electrode. Different numbers of single-stranded DNA probes are immobilized on the plurality of working electrodes.

  On the other hand, the nucleic acid concentration quantitative analysis apparatus includes a plurality of probe pins that contact a plurality of measurement electrodes, detection means, analysis means, and the like. The detection means includes a voltage application circuit that is connected to the probe pin and applies a voltage to the counter electrode, and a current detection circuit that detects a current from the working electrode.

  This system determines the analyte concentration as follows. First, a single-stranded sample DNA and a single-stranded DNA probe are subjected to a hybridization reaction (double-stranded reaction) to bind a nucleic acid insertion agent. Thereafter, a plurality of probe pins are brought into contact with the plurality of measurement electrodes, and a voltage is applied to the plurality of counter electrodes by the detection means, and a plurality of minute reaction currents from the plurality of working electrodes are detected. Thereafter, the concentration of the specimen is obtained by the analyzing means based on the detected plurality of current values (see, for example, Patent Documents 1 to 3).

JP 2004-309462 A Japanese Patent Laid-Open No. 2006-266795 JP 2009-244186 A

  Before obtaining the analyte concentration using the above-described system, it is desirable to conduct an energization test of the nucleic acid concentration quantitative analyzer, particularly an energization test of a current detection circuit responsible for detecting a minute current.

  As a method for conducting a current detection of the current detection circuit, a reaction current is detected using a sample DNA having a known concentration, and the current value is compared with a current value to be obtained. There is a method of comparing the concentration. However, when this method is used, it is necessary to supply the sample DNA to the working electrode to cause a hybridization reaction, and therefore, it takes a considerable time for the energization test. In addition, since the DNA chip cannot be reused, the cost is increased. Furthermore, since the same concentration of sample DNA is supplied to a plurality of working electrodes, the same current value may be detected from different working electrodes, and it is not possible to inspect even individual signal arrangements of the current detection circuit.

  In addition, a method for testing using an electrical measuring instrument such as a tester is conceivable, but it is necessary to disassemble the nucleic acid concentration quantitative analyzer, which takes time and man-hours.

  Therefore, the present invention has been made to solve the above-described problems, and an object of the present invention is to accurately conduct a current test of a current detection type nucleic acid concentration quantitative analysis apparatus at a low cost in a short time.

  In order to achieve the above-mentioned object, a current-carrying test cassette according to the present invention includes a plurality of working electrodes, counter electrodes, and a plurality of the plurality of working electrodes to which a single-stranded DNA probe that hybridizes with a single-stranded sample DNA is immobilized. A plurality of first current detection pads connected to the working electrode; a DNA chip on which the first voltage application pad connected to the counter electrode is formed; and the plurality of first current detections housed in the DNA chip A holder capable of mounting an analysis cassette having a pad and a chip container having a first opening formed at a position corresponding to the first voltage application pad; and the plurality of first current detections inserted through the first opening. A probe pin abutting on the pad and the first voltage application pad; and a plurality of reaction currents connected to the probe pin to apply a voltage to the counter electrode and from the plurality of working electrodes A cassette for energization testing used for an energization test of a current detection type nucleic acid concentration quantitative analysis apparatus, comprising: a detection means for detecting a concentration; and an analysis means for obtaining a concentration of a sample based on a plurality of current values detected by the detection means. A plurality of second current detection pads, a plurality of resistance parts connected to the plurality of second current detection pads and having different resistance values, and a second voltage application connected to the plurality of resistance parts A resistive circuit board on which a pad is formed, and a board that accommodates the resistive circuit board and has a second opening formed at a position corresponding to the plurality of second current detection pads and the voltage application pad and can be attached to the holder And a container.

  In order to achieve the above-described object, the energization test method according to the present invention includes a plurality of working electrodes, counter electrodes, and the like, on which a single-stranded DNA probe that hybridizes with a single-stranded sample DNA is immobilized. A plurality of first current detection pads connected to a plurality of working electrodes; a DNA chip on which a first voltage application pad connected to the counter electrode is formed; and the plurality of first currents housed in the DNA chip. A holder capable of mounting an analysis cassette having a detection pad and a chip container having a first opening formed at a position corresponding to the first voltage application pad; and the plurality of first currents inserted through the first opening A probe pin that contacts the detection pad and the first voltage application pad, and a plurality of reaction currents connected to the probe pin to apply a voltage to the counter electrode and from the plurality of working electrodes A current detecting type nucleic acid concentration quantitative analysis apparatus comprising: a detecting means for detecting a concentration; and an analyzing means for obtaining a sample concentration based on a plurality of current values detected by the detecting means. The second current detection pad, the plurality of resistance portions connected to the plurality of second current detection pads and having different resistance values, and the second voltage application pad connected to the plurality of resistance portions are formed. A current-carrying inspection cassette comprising: a resistance circuit board; and a substrate container that houses the resistance circuit board and has a second opening formed at a position corresponding to the plurality of second current detection pads and the voltage application pad. After the first step of attaching to the nucleic acid concentration quantitative analysis device and the first step, the probe pin is inserted from the second opening and is applied to the plurality of second current detection pads and the second voltage application pads. After the second step, after the second step, by applying a voltage to the plurality of resistance portions to detect a plurality of current values from the plurality of second current detection pads, and after the third step And a fourth step of comparing the resistance values of the plurality of resistance portions calculated based on the plurality of current values detected by the detection means with the actual resistance values of the plurality of resistance portions. And

  According to the present invention, it is possible to accurately conduct a current test of a current detection type nucleic acid concentration quantitative analysis apparatus at a low cost in a short time.

It is a top view of a DNA chip. It is a perspective view of the cassette for analysis. FIG. 3 is an exploded perspective view of FIG. 2. It is a perspective view of the flow path packing in FIG. It is the schematic inside the housing | casing of an analyzer. It is a schematic block diagram of an analyzer. It is a perspective view of the cassette for electric current inspection. It is a disassembled perspective view of the cassette for electricity inspection. It is a top view of a resistance circuit board. It is a circuit diagram of the resistance circuit of a resistance circuit board.

[First Embodiment]
An energization inspection cassette and an energization inspection method according to the first embodiment of the present invention will be described.

  First, a current detection type nucleic acid concentration quantitative analysis system will be described with reference to FIGS.

  A current detection type nucleic acid concentration quantitative analysis system (hereinafter simply referred to as “analysis system”) includes a nucleic acid concentration quantitative analysis cassette (hereinafter simply referred to as “analysis cassette”) 30 in which a DNA chip 10 is incorporated. , A nucleic acid concentration quantitative analyzer (hereinafter simply referred to as “analyzer”) 60 to which the analysis cassette 30 is attached. The analysis system detects and analyzes a plurality of reaction currents from the DNA chip 10 to obtain the concentration of the sample DNA.

  The DNA chip 10 will be described with reference to FIG. FIG. 1 is a top view of the DNA chip 10.

  The DNA chip 10 includes a rectangular plate-shaped insulating substrate, and a metal layer and an insulating layer stacked on the insulating substrate. The insulating substrate and the insulating layer are made of, for example, glass, Si, resin, or the like. The metal layer is made of, for example, gold. The metal layer and the insulating layer are patterned, and a plurality of three-electrode systems 12 (a plurality of working electrodes 13, a plurality of counter electrodes 14, and a plurality of reference electrodes 15), a plurality of measuring electrodes are formed on the main surface of the insulating substrate. 18 and a plurality of wirings that electrically connect the three types of electrodes 13 to 15 of the three-electrode system 12 and the measurement electrode 18 are formed.

  The three electrodes 13 to 15 and the measurement electrode 18 of the three-electrode system 12 are exposed on the main surface of the DNA chip 10. The portions other than the three types of electrodes 13 to 15 and the measurement electrode 18 of the three-electrode system 12 on the main surface of the DNA chip 10 are covered with an insulating layer.

  As shown in FIG. 1, the plurality of three-electrode systems 12 are arranged in one region (the upper region in FIG. 1) in the major axis direction on the main surface of the DNA chip 10. In the present embodiment, for example, 30 three-electrode systems 12 are arranged in 5 rows and 6 columns. Each three-electrode system 12 has two working electrodes 13, one counter electrode 14, and one reference electrode 15. The two working electrodes 13 are arranged between the counter electrode 14 and the reference electrode 15.

  Further, the plurality of measurement electrodes 18 are arranged in the other region in the major axis direction on the main surface of the DNA chip 10 (the lower region in FIG. 1). In the present embodiment, 72 measurement poles 18 are arranged in 4 rows and 18 columns. The 60 measuring electrodes 18 are electrically connected to 60 working electrodes 13 via wirings (the measuring electrodes 18 connected to the working electrodes 13 are referred to as “current detection pads 18a”). . In addition, the two measurement electrodes 18 are electrically connected to 30 counter electrodes 14 through a wiring as a common electrode (the measurement electrode 18 connected to the counter electrode 14 is referred to as a “voltage application pad 18b”. Call it.) Further, the two measurement electrodes 18 are electrically connected to the 30 reference electrodes 15 through a wiring as a common electrode.

  The working electrode 13 is an electrode on which a single-stranded DNA probe is immobilized, and is an electrode for detecting a reaction current. In this embodiment, for example, the surface areas of 60 working electrodes 13 are different from each other, and different numbers of single-stranded DNA probes proportional to the respective surface areas are fixed to the 60 working electrodes 13. ing.

  The counter electrode 14 is an electrode that supplies a current by applying a predetermined voltage to the working electrode 13. The reference electrode 15 is an electrode that feeds back an electrode voltage to the counter electrode 14 in order to control the voltage between the working electrode 13 and a predetermined voltage characteristic. By this feedback, the voltage applied by the counter electrode 14 is controlled, and highly accurate current detection that is not affected by various detection conditions can be performed.

  The analysis cassette 30 will be described with reference to FIGS. FIG. 2 is a perspective view of the analysis cassette 30. FIG. 3 is an exploded perspective view of FIG. FIG. 4 is a perspective view of the flow path packing 50 in FIG.

  The analysis cassette 30 includes a DNA chip 10, a chip container 40, and a flow path packing 50. The DNA chip 10 and the flow path packing 50 are accommodated in the chip container 40.

  The chip container 40 is formed in a rectangular box shape. The chip container 40 includes a top cover 42 and a bottom cover 44. The bottom cover 44 is screwed to the top cover 42.

  The top cover 42 is formed with injection holes 42a and discharge holes 42b for specimens, electrolytic solutions, and chemical solutions. The top cover 42 has a measurement opening 42c into which a plurality of probe pins 82 are inserted. The measurement openings 42 c are formed at positions corresponding to the plurality of measurement electrodes 18 of the DNA chip 10. In addition, arrangement | positioning of the measurement opening 42c was shown with the broken line 142c on the main surface of the DNA chip 10 of FIG.

  The DNA chip 10 is placed on the bottom cover 44 so that the main surface on which a plurality of electrodes are formed faces upward. The DNA chip 10 is disposed between the top cover 42 and the bottom cover 44.

  The flow path packing 50 is made of resin and is formed in a substantially plate shape. A plurality of three-electrode systems 12 are mounted on a region on the main surface of the DNA chip 10 formed thereon. When the bottom cover 44 is screwed to the top cover 42, the flow path packing 50 is sandwiched between the DNA chip 10 and the top cover 42 and is in close contact with the main surface of the DNA chip 10.

  A serpentine groove 52 is formed on the lower surface of the flow path packing 50 (contact surface with the DNA chip 10). The groove 52 is formed at a position corresponding to the plurality of three-electrode systems 12. In addition, two protrusions 54a and 54b are formed on the upper surface of the flow path packing 50 (the back surface of the contact surface with the DNA chip 10). The protrusions 54a and 54b are formed at positions corresponding to the injection hole 42a and the discharge hole 42b of the top cover 42. In the flow path packing 50, through holes 56a and 56b extending from the end of the groove 52 through the inside of the protrusions 54a and 54b to the injection hole 42a and the discharge hole 42b are formed. The groove 52 and the through holes 56a and 56b serve as a channel 58 for a specimen, an electrolytic solution, a chemical solution, and the like. The arrangement of the flow path packing 50 and the flow path 58 is indicated by broken lines 150 and 158 on the main surface of the DNA chip 10 in FIG.

  The analyzer 60 will be described with reference to FIGS. 5 and 6. FIG. 5 is a schematic diagram of the inside of the housing 62 of the analyzer 60. FIG. 6 is a schematic block diagram of the analyzer 60.

  The analysis apparatus 60 is equipped with the analysis cassette 30, detects and analyzes a plurality of reaction currents from the DNA chip 10, and obtains the concentration of the sample DNA. The analysis apparatus 60 includes a cassette support means 70, a probe pin unit 80, a probe pin drive means 86, a detection means 90, an analysis means 100, and the like inside a housing 62.

  The cassette support means 70 includes a cassette holder 72, a guide rail 74, a cassette holder driving means 76, and the like.

  The cassette holder 72 is formed in a substantially rectangular parallelepiped shape, and a substantially rectangular support recess 72 a corresponding to the outer shape of the analysis cassette 30 is formed in the approximate center of the upper surface thereof. The analysis cassette 30 is mounted in the support recess 72 a of the cassette holder 72.

  The guide rail 74 supports the cassette holder 72 so as to be horizontally movable. The holder driving means 76 moves the cassette holder 72 between a mounting position where the analysis cassette 30 can be removed (broken line in FIG. 5) and a measuring position where the DNA chip 10 can be measured (solid line in FIG. 5). Let The holder driving means 76 includes a plurality of driving rollers, a motor that drives the driving rollers, and the like. The casing 62 is formed with a cassette insertion port 62a through which the cassette holder 72 passes.

  The probe pin unit 80 has a plurality of probe pins 82 and a probe pin holder 84. The plurality of probe pins 82 are held by a probe pin holder 84 with the pin tips facing downward. The plurality of probe pins 82 are arranged at intervals from each other in accordance with the arrangement of the measurement electrodes 18 of the DNA chip 10. In the present embodiment, the number of probe pins 82 is 72, which is the same as the number of measurement poles 18. The 72 probe pins 82 are arranged in 4 rows and 18 columns, similarly to the arrangement of the measurement poles 18.

  The probe pin driving means 86 has a column 86a, a support arm 86b, and an arm driving means 86c. The column 86 a is erected on the bottom plate of the housing 62. The support arm 86b extends horizontally and is supported by the column 86a. The support arm 86b is moved in the horizontal direction and the vertical direction by the arm driving means 86c. The probe holder 84 is attached to the support arm 86b, and moves up and down and horizontally with the support arm 86b.

  When detecting the reaction current, the probe pin driving means 86 moves the probe pin unit 80 in the horizontal direction and the vertical direction, and passes the plurality of probe pins 82 through the measurement openings 42c of the analysis cassette 30 at the measurement position. It is brought into contact with the measuring electrode 18 of the chip 10.

  The detection means 90 includes a detection application circuit board 92 having a current detection circuit 92a for detecting a reaction current and a voltage application circuit 92b for applying a voltage. The detection application circuit board 92 is electrically connected to a plurality of probe pins 82. The voltage application circuit 92 b of the detection application circuit board 92 applies a voltage to the measurement electrode 18 (voltage application pad 18 b) connected to the counter electrode 14 via the wiring via the probe pin 82. At the same time, the current detection circuit 92a of the detection application circuit board 92 detects the reaction current from the measurement electrode 18 (current detection pad 18a) connected to the working electrode 13 by wiring via the probe pin 82.

  For example, the voltage application circuit 92b of the detection application circuit board 92 applies a voltage of 1.2 V or less, and the current detection circuit 92a of the detection application circuit board 92 can detect a minute current of 300 nA or less. The accuracy of current detection of the current detection circuit 92a is less than 1%.

  The interface unit 120 is connected to the detection application circuit board 92. The interface unit 120 is connected to an arithmetic processing unit 102 described later. The interface unit 120 inputs a signal based on the reaction current from the detection application circuit board 92 and outputs the signal to the arithmetic processing unit 102.

  The analysis unit 100 includes an arithmetic processing unit 102 and a storage unit 104. The arithmetic processing unit 102 receives a signal from the interface unit 120, reads an analysis program stored in the storage unit 104, and obtains the concentration of the sample DNA. The obtained concentration data, detected current value data, and the like are stored in the storage unit 104.

  In addition, although not shown in FIG. 5, the analyzer 60 includes a liquid feeding means 126 for supplying a sample, an electrolytic solution, a chemical solution, and the like to the DNA chip 10 inside the housing 62, and a temperature for adjusting the temperature of the DNA chip 10. The adjusting means 128 and the control part 124 which controls these are provided. The control unit 124 is connected to the interface unit 120. The control unit 124 controls the probe pin driving unit 86, the liquid feeding unit 126, and the temperature adjusting unit 128 based on a control program stored in the storage unit 104 and a user input.

  In addition, the analysis device 60 includes a user interface 122 such as a keyboard, a mouse, and a monitor outside the housing 62. The user interface 122 is connected to the arithmetic processing unit 102.

  Next, a current detection type nucleic acid concentration quantitative analysis method (hereinafter simply referred to as “analysis method”) will be described.

  An analysis cassette 30 having a DNA chip 10 in which a DNA probe is immobilized on a plurality of working electrodes 13 having different surface areas is prepared. A single-stranded DNA probe that hybridizes with a single strand of DNA whose concentration is to be examined is immobilized. The analysis cassette 30 is mounted on the cassette holder 72 of the analyzer 60 and moved to the measurement position.

  Subsequently, the single-stranded sample DNA is injected into the flow path 58 of the flow path packing 50 from the injection hole 42 a by the liquid feeding means 126 and supplied to the plurality of working electrodes 13. In addition, the temperature control means 128 heats the DNA chip 10 through the opening of the chip container 40 to cause a hybridization reaction between the DNA probe and the sample DNA. Subsequently, the liquid feeding means 126 injects a washing liquid into the flow path 58 to wash away the single-stranded sample DNA remaining without reacting. Thereafter, the nucleic acid insertion agent is injected into the flow path 58 by the liquid feeding means 126 and supplied to the working electrode 13, the counter electrode 14 and the reference electrode 15. The nucleic acid insertion agent has electrochemically active properties and selectively binds to double-stranded DNA.

  In this state, the probe pin unit 80 is moved to bring the tips of the plurality of probe pins 82 into contact with the measurement electrode 18 of the DNA chip 10. With the plurality of probe pins 82 in contact with the plurality of measurement electrodes 18, the voltage application circuit 92 b of the detection application circuit board 92 is connected between the plurality of sets of the working electrode 13 and the counter electrode 14 (the voltage application pad 18 b and the current). A predetermined voltage is applied to the detection pad 18a), and a reaction current flowing through the plurality of working electrodes 13 is detected by the current detection circuit 92a of the detection application circuit board 92 using the three-terminal method. Based on the plurality of reaction currents detected by the detection means 90, the analysis means 100 obtains the concentration of the sample DNA.

  In the present embodiment, the surface areas of the plurality of working electrodes 13 are different from each other, and different numbers of DNA probes proportional to the respective surface areas are fixed to the plurality of working electrodes 13. Therefore, the larger the surface area of the working electrode 13, the more double strands exist and the reaction current increases. The analysis unit 100 obtains calibration data from a plurality of current values and obtains the concentration based on the calibration data.

  Next, the electricity supply inspection cassette and the electricity supply inspection method according to the present embodiment will be described.

  A current inspection cassette 230 according to the present embodiment will be described with reference to FIGS. FIG. 7 is a perspective view of the power supply inspection cassette 230. FIG. 8 is an exploded perspective view of the cassette 230 for energization inspection. FIG. 9 is a top view of the resistance circuit board 210. FIG. 10 is a circuit diagram of the resistance circuit 220 of the resistance circuit board 210.

  The electricity inspection cassette 230 is mounted on the above-described analyzer 60 and is mainly used for the electricity inspection of the current detection circuit 92a of the detection application circuit board 92. The electricity inspection cassette 230 includes a resistance circuit board 210 and a board container 240. The resistance circuit board 210 is accommodated in the board container 240.

  The resistance circuit board 210 is formed to have the same outer shape and dimensions as the DNA chip 10. The resistance circuit board 210 includes an insulating substrate, a metal layer stacked on the insulating substrate, a resistance layer, and an insulating layer. The metal layer, the resistance layer, and the insulating layer are patterned, and the measurement electrode 218 (voltage application pad 218a) is provided on the main surface of the insulating substrate via the plurality of measurement electrodes 218, the plurality of resistance portions 222, and the resistance portion 222. ) And the measurement electrode 218 (current detection pad 218b) are formed with a plurality of wirings 224.

  The plurality of measurement electrodes 218 are exposed on the main surface of the resistance circuit board 210. Note that portions other than the measurement electrode 218 on the main surface of the resistance circuit board 210 are covered with an insulating layer.

  In the present embodiment, as shown in FIG. 10, 72 measurement poles 218 of the resistance circuit board 210 are formed in the same number as the measurement poles 18 of the DNA chip 10 at positions corresponding to the measurement poles 18 of the DNA chip 10. ing. Of the 72 measurement electrodes 218, the current detection pads 218a of the resistance circuit board 210 are formed in the same number as the current detection pads 18a of the DNA chip 10 at 60 positions at positions corresponding to the current detection pads 18a of the DNA chip 10. Yes. Further, two voltage application pads 218b of the resistive circuit board 210 are formed at the positions corresponding to the voltage application pads 18b of the DNA chip 10 in the same number as the voltage application pads 18b of the DNA chip 10. One resistor 222 is connected to each current detection pad 218a of the resistor circuit board 210. A voltage is applied to the 60 resistor portions 222 from the voltage application pad 218b which is a common electrode.

  The plurality of resistance units 222 have different resistance values. The resistance value of the resistance unit 222 is set to be equal to the resistance value of the DNA chip 10, that is, the resistance value between the counter electrode 14 and the working electrode 13. The resistance value of the resistance unit 222 to be set includes the voltage applied by the detection application circuit board 92 to the DNA chip 10 (for example, 1.2 V or less) and the current detected by the detection application circuit board 92 (for example, 300 nA or less). ) Based on Ohm's law. The resistance values of the plurality of resistor units 222 are set to, for example, several M ohms or more. Since the detection accuracy of the detection application circuit board 92 is less than 1%, it is desirable to set different resistance values for the plurality of resistance portions 222 by 1% or more.

  The substrate container 240 includes a top cover 242 and a bottom cover 244. The outer shape of the substrate container 240 is formed to have the same dimensions as the outer shape of the chip container 40. A measurement opening 242 c is formed in the top cover 242 of the substrate container 240 at the same position as the top cover 42 of the chip container 40. In addition, arrangement | positioning of the measurement opening 242c was shown with the broken line 442c on the main surface of the resistive circuit board 210 of FIG. In the present embodiment, unlike the top cover 42 of the chip container 40, the injection hole and the discharge hole are not formed in the top cover 242 of the substrate container 240.

  The resistance circuit board 210 is placed on the bottom cover 244 so that the main surface on which the plurality of measurement electrodes 218 are formed faces upward. The resistance circuit board 210 is disposed between the top cover 242 and the bottom cover 244 and is fixed in the board container 240. In the present embodiment, the electricity inspection cassette 230 does not have a flow path packing.

  Hereinafter, the energization inspection method according to the present embodiment will be described.

  First, the electricity inspection cassette 230 is mounted on the cassette holder 72 of the analyzer 60 and moved to the measurement position. Since the substrate container 240 is formed in the same shape as the outer shape of the chip container 40, the substrate container 240 is attached to the support recess 72 a of the cassette holder 72.

  In this state, the probe pin unit 80 is moved, and the tips of the plurality of probe pins 82 are brought into contact with the measurement electrode 218 of the resistance circuit board 210. In a state where the plurality of probe pins 82 are in contact with the plurality of measurement electrodes 218, the voltage application circuit 92b of the detection application circuit board 92 causes a plurality of sets of measurement electrodes 218 (between the voltage application pad 218a and the current detection pad 218b). In addition, a predetermined voltage is applied, and a reaction current flowing through the plurality of resistance portions 222 is detected by the current detection circuit 92a of the detection application circuit board 92 using the three-terminal method. By comparing a plurality of resistance values calculated based on the applied voltage and the detected current and a plurality of actual resistance values of the resistor section 222, energization confirmation of the current detection circuit 92a of the detection application circuit board 92 is performed. .

  Hereinafter, the effects of the electricity inspection cassette 230 and the electricity inspection method according to the present embodiment will be described.

  As described above, before obtaining the concentration of the sample using the analysis system, it is desirable to conduct an energization test of the nucleic acid concentration quantitative analyzer 60, particularly an energization test of the current detection circuit 92a responsible for detecting a minute current. .

  According to the present embodiment, since the current inspection cassette 230 is used, there is no need to supply the sample DNA to the working electrode 13 to cause a hybridization reaction, and the current detection circuit 92a of the detection application circuit board 92 is energized in a short time. Inspection can be performed. In addition, the energization inspection cassette 230 can be reused, and the cost for the energization inspection can be suppressed. Further, a plurality of resistance circuits 220 having different resistance values are formed in the current-carrying inspection cassette 230. In order to conduct a current-carrying inspection by detecting currents flowing through these resistance circuits 220, a current detection circuit 92a. Up to individual signal configurations.

  Furthermore, the energization check between the probe pin 82 and the measurement electrode 18 or the energization confirmation between the probe pin 82 and the detection application circuit board 92 may be performed using the energization inspection method according to the present embodiment.

[Other Embodiments]
The above embodiment is merely an example, and the present invention is not limited to the above embodiment. For example, in the above embodiment, the resistance circuit 220 is formed by patterning a metal layer, a resistance layer, and an insulating layer stacked on an insulating substrate, but the metal layer stacked on the insulating substrate is patterned. The wiring 224 formed in this way may be formed by soldering a resistance chip.

  In the above embodiment, the top cover 242 of the substrate container 240 is not formed with the injection hole 42a and the discharge hole 42b that are not used in the energization inspection, but these may be formed. Since the chip container 40 is used as the substrate container 240, the manufacturing cost can be reduced.

DESCRIPTION OF SYMBOLS 10 ... DNA chip, 12 ... 3 electrode system, 13 ... Working electrode, 14 ... Counter electrode, 15 ... Reference electrode, 18 ... Measuring electrode, 18a ... Current detection pad, 18b ... Voltage application pad, 30 ... Analysis cassette, 40 ... Chip container, 42 ... top cover, 42a ... injection hole, 42b ... discharge hole, 42c ... measurement opening, 44 ... bottom cover, 50 ... flow path packing, 52 ... groove, 54a, 54b ... projection, 56a, 56b ... penetration Hole: 60 ... Analyzer, 62 ... Case, 62a ... Cassette insertion port, 70 ... Cassette support means, 72 ... Cassette holder, 72a ... Support recess, 74 ... Guide rail, 76 ... Holder drive means, 80 ... Probe pin unit , 82 ... Probe pin, 84 ... Probe pin holder, 86 ... Probe pin driving means, 90 ... Detection means, 92 ... Detection application circuit board, 92a ... Current detection circuit 92b ... Voltage application circuit, 100 ... analyzing unit, 102 ... calculation processing unit, 104 ... storage unit, 120 ... interface unit, 122 ... user interface, 124 ... control unit, 126 ... liquid feeding unit, 128 ... temperature adjusting unit, 210 ... resistance circuit board, 218 ... measurement electrode, 218a ... current detection pad, 218b ... voltage application pad, 220 ... resistance circuit, 222 ... resistance part, 224 ... wiring, 230 ... cassette for current inspection, 240 ... substrate container, 242 ... Top cover, 242c ... Measurement opening, 244 ... Bottom cover

Claims (8)

A plurality of working electrodes, a counter electrode, and a plurality of first current detection pads connected to the plurality of working electrodes, to which a single-stranded DNA probe for hybridizing a single-stranded sample DNA is immobilized, and the counter electrode A DNA chip on which a first voltage application pad is formed, and a first opening is formed at a position corresponding to the plurality of first current detection pads and the first voltage application pad. A holder capable of mounting an analysis cassette having a chip container,
Probe pins inserted from the first openings and contacting the first current detection pads and the first voltage application pads;
Detecting means connected to the probe pin and applying a voltage to the counter electrode to detect a plurality of reaction current values from the plurality of working electrodes;
Analyzing means for obtaining the concentration of the specimen based on a plurality of current values detected by the detecting means;
A cassette for energization inspection used for energization inspection of a current detection type nucleic acid concentration quantitative analysis apparatus comprising:
A plurality of second current detection pads, a plurality of resistance parts connected to the plurality of second current detection pads and having different resistance values, and a second voltage application pad connected to the plurality of resistance parts are formed. A resistive circuit board,
A substrate container accommodating the resistor circuit board and having a second opening formed at a position corresponding to the plurality of second current detection pads and the voltage application pad;
A cassette for energization inspection characterized by comprising:   2. The cassette for energization inspection according to claim 1, wherein the number of the second current detection pads is the same as the number of the first current detection pads.   The number of the said 2nd voltage application pad is the same number as the number of the said 1st voltage application pad, The cassette for electricity supply inspection of Claim 1 or 2 characterized by the above-mentioned.   The external inspection cassette according to any one of claims 1 to 3, wherein the outer shape of the substrate container has the same dimensions as the outer shape of the chip container.   The cassette for energization inspection according to claim 4, wherein a position where the second opening of the substrate container is formed is the same position as a position where the first opening of the chip container is formed.   6. The cassette for energization inspection according to claim 1, wherein the plurality of resistance portions have resistance values different from each other more than a current detection accuracy of the detection unit. A plurality of working electrodes, a counter electrode, and a plurality of first current detection pads connected to the plurality of working electrodes, to which a single-stranded DNA probe for hybridizing a single-stranded sample DNA is immobilized, and the counter electrode A DNA chip on which a first voltage application pad is formed, and a first opening is formed at a position corresponding to the plurality of first current detection pads and the first voltage application pad. A holder capable of mounting an analysis cassette having a chip container,
Probe pins inserted from the first openings and contacting the first current detection pads and the first voltage application pads;
Detecting means connected to the probe pin and applying a voltage to the counter electrode to detect a plurality of reaction current values from the plurality of working electrodes;
Analyzing means for obtaining the concentration of the specimen based on a plurality of current values detected by the detecting means;
A current detection type nucleic acid concentration quantitative analysis apparatus comprising:
A plurality of second current detection pads, a plurality of resistance parts connected to the plurality of second current detection pads and having different resistance values, and a second voltage application pad connected to the plurality of resistance parts are formed. A resistive circuit board,
A substrate container in which a second opening is formed at a position corresponding to the plurality of second current detection pads and the voltage application pad, containing the resistance circuit board;
A first step of mounting a cassette for energization inspection having the nucleic acid concentration quantitative analysis apparatus,
After the first step, a second step in which the probe pin is inserted from the second opening and brought into contact with the plurality of second current detection pads and the second voltage application pad;
A third step of detecting a plurality of current values from the plurality of second current detection pads by applying a voltage to the plurality of resistance portions after the second step;
A fourth step of comparing the resistance values of the plurality of resistance portions calculated based on the plurality of current values detected by the detection means with the actual resistance values of the plurality of resistance portions after the third step;
An electrical inspection method characterized by comprising:   The energization inspection method according to claim 7, wherein the detection unit includes a current detection circuit that detects a current value, and inspects the energization of the current detection circuit.

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