JP2009150809A - Microchip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microchip capable of detecting existence of a specific object material by generating an amplification reaction, and detecting accurately an influence of inhibition of amplification. <P>SOLUTION: This microchip has a constitution having a specimen dividing part SP for dividing specimen liquid sent through a fine channel from a specimen storage part stE into a first channel and a second channel. In the fine channel, the first mixed liquid is formed by joining together the specimen liquid sent through the first channel, positive control from a positive control storage part stP, and a reagent from a reagent storage part st, and the second mixed liquid is formed by joining together the specimen liquid sent through the second channel, negative control from a negative control storage part stN, and the reagent from the reagent storage part st, and the first mixed liquid and the second mixed liquid are amplified respectively by a nucleic acid amplification reaction, and the amplification reaction is detected by a detection part based on existence of amplification. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a microchip for genetic testing.

  In recent years, by making full use of micromachine technology and ultrafine processing technology, devices and means (for example, pumps, valves, flow paths, sensors, etc.) for performing conventional sample preparation, chemical analysis, chemical synthesis, etc. have been miniaturized. A system integrated on a chip attracts attention. This is also referred to as μ-TAS (Micro Total Analysis System), and a DNA-treated extraction solution obtained by processing a reagent and a sample (for example, urine, saliva, blood of a subject to be tested) on a member called a microchip. Etc.) and the characteristics of the specimen are examined by detecting the reaction.

  Microchips use a photolithographic process (a method of creating a groove by etching a pattern image with a chemical) or a groove using a laser beam on a substrate made of a resin material or glass material, and flow a reagent or specimen. A fine channel and a liquid reservoir for storing reagents have been provided, and various patterns have been proposed.

  When investigating the characteristics of a specimen using these microchips, the reagent and specimen contained in the microchip are sent by a micropump or the like, and the reagent and specimen are subjected to an amplification reaction to the detection section. Guide and detect. In the detection unit, the target substance is detected by, for example, an optical detection method.

  The microchip can detect even a small amount of sample by performing an amplification reaction. When such an amplification reaction is performed, a test error may occur due to contamination or contamination of reaction inhibiting substances, or failure of amplification due to reagent inactivation, inappropriate reaction conditions, or the like. It is customary to analyze the controls in parallel to detect that such an inspection error has occurred.

In Patent Document 1, a positive control and a negative control are used as controls, and a mixture of a specimen and a reagent (1) A mixture of a positive control and a reagent (2)
A liquid mixture (3) of the negative control and the reagent is obtained. And then
(A) a mixture of the mixture (1) and the mixture (2);
(B) Only mixed liquid (1),
(C) Mixture (2) only,
(D) Only the mixture (3),
A microreactor is disclosed in which each fluid is subjected to an amplification reaction in an amplification section to detect the increased production substance.
International Publication No. 07/058077 Pamphlet

  However, the microreactor disclosed in Patent Document 1 has a problem that the influence of amplification inhibition cannot be accurately detected because the ratio of the inhibitory substance brought from the specimen is different between the fluid (a) and the fluid (b). .

  In view of the above problems, an object of the present invention is to obtain a microchip capable of detecting the presence of a specific target substance by performing an amplification reaction and accurately detecting the influence of amplification inhibition.

  The above object is achieved by the invention described below.

(1) a sample container into which a sample solution containing a sample or DNA extracted from the sample is injected;
A reagent container for storing a reagent used in the nucleic acid amplification reaction;
A positive control accommodating part for accommodating a positive control;
A negative control accommodating portion for accommodating a negative control;
A fine flow path communicating with each of these accommodating portions,
A detection unit for detecting an amplification reaction;
A microchip having
A sample dividing section that divides the sample liquid fed from the sample storage section into the first flow path and the second flow path;
The fine channel is
A sample liquid fed through the first flow path, a positive control from the positive control storage unit and a reagent from the reagent storage unit are merged to form a first mixed solution;
A sample liquid fed through the second flow path, a negative control from the negative control storage unit and a reagent from the reagent storage unit are merged to form a second mixed solution;
A microchip configured to amplify a first mixed solution and a second mixed solution by a nucleic acid amplification reaction and detect the amplification reaction based on the presence or absence of amplification by the detection unit.

  (2) It has a probe accommodating portion, and is configured to detect by hybridization of a nucleic acid to be detected amplified for the presence or absence of amplification and a probe from the probe accommodating portion (1 ) Microchip.

  (3) It has a probe housing part, and is configured to detect by a temperature difference at the time of a hybridization reaction between the nucleic acid to be detected amplified for the presence or absence of amplification and the probe from the probe housing part. The microchip according to (1), which is characterized.

  (4) It has a probe storage unit, and is configured to detect the presence / absence of amplification by hybridization between a nucleic acid to be detected being amplified and a probe from the probe storage unit (1) ) Microchip.

  According to the present invention, it is possible to obtain a microchip capable of detecting the presence of a specific target substance by performing an amplification reaction and accurately detecting the influence of amplification inhibition.

  Although the present invention will be described based on an embodiment, the present invention is not limited to the embodiment.

[Analytical system configuration]
FIG. 1 is an external view of a microchip analysis system 8 using the microchip according to the present embodiment. The microchip analysis system 8 is a device that automatically reacts a sample and a reagent previously injected into the microchip 1 and automatically outputs a reaction result.

  A housing 82 of the microchip analysis system 8 includes an insertion port 83 for inserting the microchip 1 into the apparatus, a display unit 84, a memory card slot 85, a print output port 86, an operation panel 87, and an external input / output terminal 88. Is provided.

  The person in charge of inspection inserts the microchip 1 in the direction of the arrow in FIG. 1 and operates the operation panel 87 to start the inspection. In the microchip analysis system 8, the reaction in the microchip 1 is automatically inspected, and when the inspection is completed, the result is displayed on the display unit 84. The inspection result can be output from the print output port 86 or stored in a memory card inserted in the memory card slot 85 by operating the operation panel 87. Further, data can be stored in the personal computer or the like from the external input / output terminal 88 using, for example, a LAN cable. After completion of the inspection, the inspection person takes out the microchip 1 from the insertion port 83.

  FIG. 2 is a schematic perspective view of a microchip analysis system 8 using the microchip according to the present embodiment, and FIG. 3 is a configuration diagram. 2 and 3, the microchip is inserted through the insertion port 83 shown in FIG. 1 and the setting is completed.

  The microchip analysis system 8 includes a driving liquid tank 70 that stores a driving liquid L0 for feeding a sample and a reagent previously injected into the microchip 1, and a micropump 5 for supplying the driving liquid L0 to the microchip 1. The pump connection part 6 that connects the micropump 5 and the microchip 1 so that the driving liquid L0 does not leak, the temperature control unit 3 that controls the temperature of the necessary part of the microchip 1, and the temperature control so that the microchip 1 does not deviate. A chip pressing plate 2 for bringing the chip pressing plate 2 into close contact with the unit 3 and the pump connection unit 6; a pressing plate driving unit 21 for moving the chip pressing plate 2 up and down; a regulating member 22 for positioning the microchip 1 with respect to the micro pump 5 with high accuracy; A light detection unit 4 (4a and 4b) for detecting a reaction state between the specimen and the reagent in the microchip 1 is provided.

  The chip pressing plate 2 is retracted upward from the position shown in FIG. 3 in the initial state. Thereby, the microchip 1 can be inserted / removed in the direction of the arrow X, and the person inspecting inserts the microchip 1 from the insertion port 83 (see FIG. 1) until it comes into contact with the regulating member 22. Thereafter, the chip pressing plate 2 is lowered by the pressing plate driving unit 21 and comes into contact with the microchip 1, and the lower surface of the microchip 1 is in close contact with the temperature adjustment unit 3 and the pump connection unit 6.

  The temperature control unit 3 includes a Peltier element 31 and a heater 32 on the surface facing the microchip 1. When the microchip 1 is set in the microchip analysis system 8, the Peltier element 31 and the heater 32 are attached to the microchip 1. It comes to adhere closely. A part containing the reagent is cooled by the Peltier element 31 so that the reagent is not denatured, or the reaction part 139 where the specimen and the reagent react is heated by the heater 32 to promote the reaction.

  The control temperature at the time of heating in the reaction unit 139 can be changed as appropriate. By switching the control temperature, whether or not the nucleic acid to be detected is contained is detected based on the presence or absence of amplification by the hybridization reaction. Anyway.

  In the light detection unit 4 including the light emitting unit 4a and the light receiving unit 4b, the light from the light emitting unit 4a is irradiated to the microchip 1, and the light transmitted through the microchip 1 is detected by the light receiving unit 4b. The light receiving portion 4b is integrally provided inside the chip pressing plate 2. The light emitting unit 4a and the light receiving unit 4b are provided to face the detecting unit 148 of the microchip 1 shown in FIG.

  The micropump 5 is located on the pump chamber 52, the piezoelectric element 51 that changes the volume of the pump chamber 52, the first throttle channel 53 located on the microchip 1 side of the pump chamber 52, and the driving fluid tank 70 side of the pump chamber. The second throttle channel 54 is formed. The first throttle channel 53 and the second throttle channel 54 are narrow and narrow channels, and the first throttle channel 53 is longer than the second throttle channel 54.

  When the driving liquid L0 is fed in the forward direction (the direction toward the microchip 1), first, the piezoelectric element 51 is driven so that the volume of the pump chamber 52 is rapidly reduced. Then, a turbulent flow is generated in the second throttle channel 54 that is a short throttle channel, and the channel resistance in the second throttle channel 54 is relatively larger than that of the first throttle channel 53 that is a throttle channel. growing. As a result, the driving liquid L0 in the pump chamber 52 is predominantly pushed toward the first throttle channel 53 and fed. Next, the piezoelectric element 51 is driven so that the volume of the pump chamber 52 is gradually increased. Then, the driving liquid L0 flows from the first throttle channel 53 and the second throttle channel 54 as the volume in the pump chamber 52 increases. At this time, since the length of the second throttle channel 54 is shorter than that of the first throttle channel 53, the channel resistance of the second throttle channel 54 is smaller than that of the first throttle channel 53. Thus, the driving liquid L 0 flows into the pump chamber 52 predominantly from the second throttle channel 54. When the piezoelectric element 51 repeats the above operation, the driving liquid L0 is fed in the forward direction. Further, it is possible to change the liquid feeding pressure of the driving liquid L0 by changing the driving voltage to the piezoelectric element 51.

  On the other hand, when the driving liquid L0 is fed in the reverse direction (direction toward the driving liquid tank 70), first, the piezoelectric element 51 is driven so that the volume of the pump chamber 52 is gradually reduced. Then, since the length of the second throttle channel 54 is shorter than that of the first throttle channel 53, the channel resistance of the second throttle channel 54 is smaller than that of the first throttle channel 53. . As a result, the driving liquid L0 in the pump chamber 52 is predominantly pushed out toward the second throttle channel 54 and fed. Next, the piezoelectric element 51 is driven so that the volume of the pump chamber 52 is rapidly increased. Then, the driving liquid L0 flows from the first throttle channel 53 and the second throttle channel 54 as the volume in the pump chamber 52 increases. At this time, turbulent flow is generated in the second throttle channel 54, which is a short throttle channel, and the channel resistance in the second throttle channel 54 is relatively larger than that of the first throttle channel 53, which is a throttle channel. Become bigger. As a result, the driving liquid L 0 flows into the pump chamber 52 predominantly from the first throttle channel 53. When the piezoelectric element 51 repeats the above operation, the driving liquid L0 is fed in the reverse direction.

  The pump connection portion 6 is formed of a contact surface made of a resin having flexibility (elasticity, shape followability) such as polytetrafluoroethylene or silicon resin in order to ensure necessary sealing performance and prevent leakage of driving liquid. It is preferred that Such a close contact surface having flexibility may be, for example, due to the constituent substrate of the microchip itself, or a separate adhesive having flexibility attached around the flow path opening in the pump connection portion 6. It may be due to a member.

[Configuration of Microchip 1]
Next, the configuration of the microchip 1 according to the present embodiment will be described with reference to FIGS. 4 and 5. FIG. 4 is a schematic configuration diagram illustrating the relationship between the flow path elements of the microchip 1, and FIG. 5 is a schematic diagram illustrating an example of the flow path configuration of the microchip 1.

  As shown in FIG. 4, in the microchip 1 according to the present embodiment, the sample is divided into two by the sample dividing unit SP, and one of the divided samples is fed through the first flow path 131, and the positive control, the reagent, The first mixed liquid is made by joining. The other of the divided samples is fed through the second flow path 132 and joined with the negative control and the reagent to create a second mixed solution.

  The first mixed solution and the second mixed solution are amplified by the respective amplification units 139 and then hybridized with the probe DNA (or simply referred to as a probe) by the detection unit 148. Based on this reaction product, An amplification reaction is detected by the detection unit. The above is the outline of the whole.

  Here, the “probe DNA” refers to a DNA fragment that has a complementary relationship with a gene to be detected by utilizing the complementarity of DNA. It binds to the gene to be detected by hybridization.

  “Positive control” alone causes a nucleic acid amplification reaction similar to that of the detection target gene and a hybridization reaction between the probe DNA and a production reaction with a fluorescent substance described later. The sequence is a specific sequence for detecting the sample, and the portion where the primer hybridizes and the sequence between them are the same as the sample. Nucleic acids (DNA, RNA) used for control may be those described in known technical literature.

  The “negative control” alone does not cause a production reaction with a fluorescent substance. It contains all reagents other than nucleic acid (DNA, RNA), etc., and is used for checking for contamination and for background correction.

  The “dissociation temperature” is a temperature at which a nucleic acid having a double-stranded structure is broken and dissociates into single strands, and is also referred to as a denaturing temperature. The dissociation temperature tends to be high for duplexes with high homology and low for duplexes with low homology.

  FIG. 5 shows an example of the microchip 1 according to the present embodiment. In the same figure, arrangement | positioning of the microchannel and channel element in the state from which the coating substrate was removed is shown typically.

  The microchip 1 is provided with a fine channel and a channel element for mixing and reacting a liquid specimen (sample) on the microchip 1 using a hydrophobic base material in the same manner as a liquid reagent. Yes. The microchannel is formed in the order of micrometers, for example, the width w is several tens to several hundreds of micrometers, preferably 50 to 300 μm, and the height h is about 25 to 1000 μm, preferably 50 to 300 μm.

  Hereinafter, the reaction and detection steps in the microchip will be described. In the microchip 1 shown in FIG. 5, the two reaction detection flow paths are arranged substantially symmetrically (left half and right half in the figure). As in the schematic diagram shown in FIG. 4, the reaction detection channel in the left half is used for reaction detection between the specimen, the positive control, and the reagent, and the reaction detection channel in the right half is used for the specimen, the negative control, the reagent, and It is used for detection of reaction. By comparing the two reaction detections, the influence of amplification inhibition can be detected with high accuracy. Since the two reaction detection channels are symmetrically arranged and basically have the same channel configuration, only the reaction detection channel in the left half will be mainly described below.

  g (g1, g2, g6 to g8) is an upstream opening portion released from one surface of the microchip 1 to the outside. These upstream openings g are aligned with the flow path openings provided on the connection surface of the micropump 5 when the microchip 1 is overlapped and connected to the micropump 5 via the pump connection part 6. The pump 5 communicates. Then, a driving liquid L0 for feeding liquid is injected from the upstream opening.

  i is an injection hole for injecting a liquid such as a reagent or a specimen (hereinafter also simply referred to as a reagent solution), and is an opening opened to the outside from the upper surface of the microchip 1. The reagent solution is injected with the upstream opening g in the vicinity of each injection hole i opened. The injected liquid is sent through the fine channel toward the nearby upstream opening g. In the present embodiment, a part of the fine channel for storing the liquid is used as the reagent storage unit st (st1 to st8).

  A positive control is accommodated in the positive control accommodating part stP, and a negative control is accommodated in the negative control accommodating part stN. The sample storage unit stE stores a sample solution containing a sample or DNA extracted from the sample. As will be described later, probe DNA is accommodated in the reagent accommodating portion st6. In the present embodiment, the reagent storage unit st6 is used as a “probe storage unit”.

  At the time of reagent solution injection, only the upstream opening g and the injection hole i are open, and only the injection hole i is sealed after the reagent injection. Then, a liquid such as a reagent or a sample is sent through the air by the driving liquid L0 sent from the micropump 5 communicating with the reagent storage unit st and the upstream opening g.

  Reference numeral 130 denotes a mixed solution storage unit, in which reagent solutions from the reagent storage units st1 to st3 that have joined together on the upstream side are mixed.

  j (j1 to j6) is a merging section, and SP is a specimen dividing section. In the junction j, the reagent solution sent from the upstream microchannel is joined. The sample dividing unit SP divides the sample liquid sent from the upstream sample storage unit stE into two. One of the divided parts is fed through the first flow path 131 and merged with the positive control fed from the positive control storage unit stP and the reagent from the reagent storage units st41, st42, etc. 1 mixture "is created. Similarly, the other sample divided by the sample dividing unit SP is fed through the second flow path 132, and at the junction j6, the negative control and the reagent containing units st42 and st43 fed from the negative control containing unit stN. The “second mixed solution” is created by combining the reagents from the above.

  On the downstream side of the junctions j5 and j6, a mixing channel 138 is provided for mixing the first mixed solution and the second mixed solution with sufficient molecular diffusion. A reaction unit 139 is provided downstream of the mixing channel 138.

  The reaction unit 139 is a part that heats and reacts the mixed liquid of the sample and the reagent sufficiently mixed in the mixing channel 138. When the microchip 1 is set in the microchip analysis system 8, the reaction unit 139 performs microchip analysis. The heaters 32 of the system 8 are opposed to each other. By heating the reaction part 139 with the heater 32, a nucleic acid amplification reaction is performed. The amplification product amplified by the reaction unit 139 by the nucleic acid amplification reaction is sent to the detection unit 148 (148a, 148b) and detected by the light detection unit 4.

  A means for detecting the amplification product in the detection unit 148 will be described. The detection unit 148 cannot optically detect the amplification product as it is. In general, the amplification product is allowed to trap in the detection unit 148 by reacting the amplification product with a reactant carried on the flow path wall of the detection unit 148. In addition, a fluorescently labeled probe DNA is bound to the amplification product so that it can be detected optically. At least the detection part of the detection unit 148 is made of a transparent material, preferably a transparent plastic, in order to enable optical measurement.

  Here, the genetic test will be specifically described as an example.

  (1) The reagent is a biotin-modified primer, and nucleic acid amplification of the sample is performed in the reaction unit 139, and the sample after reaction in which the amplified gene is made into a single strand by denaturation treatment is sent to the detection unit 148. A biotin-affinity protein such as streptavidin (avidin, streptavidin, extraavidin, preferably streptavidin) is supported and immobilized in advance on the flow path wall of the detection unit 148. When the sample after reaction in the reaction unit 139 flows into the detection unit 148, the gene of the sample is immobilized on the channel wall of the detection unit 148 by a binding reaction between biotin affinity protein and biotin labeled with the probe DNA ( Trapped). The above-described binding reaction between the biotin affinity protein and biotin is a known avidin-biotin reaction.

Furthermore, through a step of trapping an amplification product (in this example, an amplification gene), a probe DNA fluorescently labeled with FITC (Fluorescein isothiocyanate) is passed to the detection unit 148 that traps the amplification gene. Hybridize to the gene immobilized on the channel wall. (A hybrid of an amplified gene and a fluorescently labeled probe DNA may be trapped by the detection unit.)
(2) A gold colloid solution whose surface is modified with an anti-FITC antibody that specifically binds to FITC is allowed to flow into the microchannel, and thereby the gold colloid is adsorbed to the FITC-modified probe hybridized with the gene.

  (3) The concentration of the gold colloid in the fine channel is optically measured.

  It is also possible to measure the fluorescence of the fluorescent dye FITC. However, it is necessary to consider light fading of the fluorescent dye, background noise, and the like. For this reason, in the present embodiment, as described above, a system that can finally measure with high sensitivity using visible light is employed.

  As described above, in the detection unit 148, each reagent contained in the fine channel is sequentially sent and reacts with the reactants immobilized on the detection unit 148. This order is determined in advance.

  The amplification product sent from the reaction unit 139 to the detection unit 148 starts a reaction with the reactant in the detection unit 148 (for example, an avidin-biotin reaction).

  Next, feeding of the driving liquid L0 from the upstream opening g6 is started, and the probe DNA accommodated in the reagent accommodating part st6 is fed to the downstream detecting part 148 to thereby hybridize with the reactant in the detecting part 148. Reaction takes place.

  After that, the feeding of the driving liquid L0 from the upstream opening g7 is started, and the antigen-antibody reaction is started in the detection unit 148 by feeding the dye solution (for example, PEGylated gold colloid) in the reagent storage unit st7 to the detection unit 148. Is done. After the gold colloid reacts with the amplification product, an extra gold colloid is present when detected by the detection unit 148. In order to wash away the excess gold colloid, the cleaning liquid in the reagent storage unit st8 is sent to the detection unit 148 by feeding the driving liquid L0 from the upstream opening g8.

  The amplification product sent to the detection unit 148 and subjected to a reaction for detection is detected by the light detection unit 4. The amplified product after detection is sent to the waste liquid section 160.

  The inspection is normal by comparing the detection results of the detection unit 148a that detects the amplified product generated from the first mixed solution and the detection unit 148b that detects the amplified product generated from the second mixed solution. It can be determined whether or not it has been performed.

  That is, when the target gene is contained in the positive sample, that is, the reaction product produced by any of the “first mixed solution” of the sample and the positive control and the “second mixed solution” of the sample and the negative control. Fluorescence emission also occurs from. When the target gene is not included in the negative sample, that is, the “first mixture” emits fluorescence due to the positive control reaction, whereas the “second mixture” does not react and does not emit fluorescence. . These two cases can be treated as test results in which a normal reaction has been performed.

  On the other hand, for example, when an amplification inhibitor is mixed in the specimen, no fluorescence is emitted from the reaction product of any of the “first mixed solution” and “second mixed solution”. Negative. Further, when there is no fluorescence emission in the reaction product of the “first mixed solution” and fluorescence emission is in the reaction product of the “second mixture”, abnormalities such as deactivation of the reagent contained in the microchip 1 are observed. Can be considered. In these two cases, re-examination can be promoted as an inspection result in which an abnormal reaction is performed.

  In this embodiment, the “second mixture” is diluted with a negative control, so that a test abnormality due to the influence of amplification inhibition is detected with high accuracy in comparison with the “first mixture”. It becomes possible to do.

It is an external view of the microchip analysis system 8 using the microchip which concerns on this embodiment. It is a schematic perspective view of the microchip analysis system 8 using the microchip which concerns on this embodiment. It is a block diagram of the microchip analysis system 8 using the microchip which concerns on this embodiment. 2 is a schematic configuration diagram illustrating a relationship between flow path elements of a microchip 1. FIG. 1 is a schematic diagram illustrating an example of a flow path configuration of a microchip 1. FIG.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microchip 6 Pump connection part g Upstream opening part st1-st8 Reagent storage part stE Specimen storage part stP Positive control storage part stN Negative control storage part i Injection hole j Confluence part SP Specimen division part 130 Mixed liquid storage part 138 Mixing channel 139 Reaction section 148 Detection section 160 Waste liquid section 70 Drive liquid tank

Claims (4)

A sample container into which a sample solution containing a sample or DNA extracted from the sample is injected;
A reagent container for storing a reagent used in the nucleic acid amplification reaction;
A positive control accommodating part for accommodating a positive control;
A negative control accommodating portion for accommodating a negative control;
A fine flow path communicating with each of these accommodating portions,
A detection unit for detecting an amplification reaction;
A microchip having
A sample dividing section that divides the sample liquid fed from the sample storage section into the first flow path and the second flow path;
The fine channel is
A sample liquid fed through the first flow path, a positive control from the positive control storage unit and a reagent from the reagent storage unit are merged to form a first mixed solution;
A sample liquid fed through the second flow path, a negative control from the negative control storage unit and a reagent from the reagent storage unit are merged to form a second mixed solution;
A microchip configured to amplify a first mixed solution and a second mixed solution by a nucleic acid amplification reaction and detect the amplification reaction based on the presence or absence of amplification by the detection unit. Having a probe housing,
2. The microchip according to claim 1, wherein the microchip is configured to be detected by hybridization of a nucleic acid to be detected that has been amplified based on the presence or absence of amplification, and a probe from the probe storage unit. Having a probe housing,
2. The micro of claim 1, wherein the detection is performed based on a temperature difference at the time of a hybridization reaction between the nucleic acid to be detected that has been amplified based on the presence or absence of amplification and the probe from the probe storage unit. Chip. Having a probe housing,
2. The microchip according to claim 1, wherein the presence or absence of amplification is detected by hybridization between a nucleic acid to be detected being amplified and a probe from the probe housing portion.

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