JP2009222479A - Inspection device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inspection device capable of performing the amplification of a gene by a PCR method using a small-sized microinspection chip. <P>SOLUTION: A liquid sending part for sending a liquid within the microinspection chip is provided, and a temperature variable part having a plurality of temperature regions and the microinspection chip are relatively moved to repeat a temperature cycle in a state that the liquid is stopped within the microinspection chip. Accordingly, the inspection device is provided, wherein the amplification of the gene by the PCR method can be performed using the small-sized microinspection chip without requiring a long flow channel on the microinspection chip. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an inspection apparatus, and in particular, microinspection used for biological substance inspection / analysis by gene amplification reaction, antigen-antibody reaction, etc., inspection / analysis of other chemical substances, chemical synthesis of target compounds by organic synthesis, etc. The present invention relates to an inspection apparatus using a chip.

  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. An analysis chip (hereinafter referred to as a micro inspection chip) integrated on a chip has been developed (see, for example, Patent Document 1).

  This is also called μ-TAS (Micro Total Analysis System), bioreactor, Lab-on-chip, biochip, and it is used in the medical examination / diagnosis field, environmental measurement field, and agricultural production field. Application is expected. In particular, when complicated processes such as those found in genetic testing, skilled techniques, and operation of equipment are required, micro test chips that excel in automation, speed-up, and simplification are cost-effective and require sample volume. Because it enables analysis not only for the required time but also for any time and place, the benefits are great.

  The present applicant injects a specimen such as blood into a micro test chip enclosing a reagent, etc., and injects a driving liquid with a micro pump into the micro flow path of the micro test chip, thereby moving the specimen and reacting sequentially. Have proposed a reaction detection device capable of measuring the results (see, for example, Patent Document 2).

  A polymerase chain reaction method (hereinafter referred to as a PCR method) is known as a gene amplification method for gene diagnosis using a micro test chip. In the PCR method, a specimen containing a gene to be amplified is amplified under a plurality of temperature conditions (for example, three temperature conditions of about 95 ° C. heat denaturation temperature, about 55 ° C. annealing temperature, and about 70 ° C. polymerization temperature). The gene can be amplified in large quantities by repeating this cycle many times.

  For example, in Patent Document 3, (1) a method for performing gene amplification by PCR by performing a temperature cycle by sequentially immersing a capillary tube containing a specimen containing a gene in a temperature bath under three temperature conditions, (2) 1 A method of performing gene amplification by the PCR method by bending a capillary tube and sequentially immersing it in a temperature bath of three temperature conditions, and flowing a sample containing the gene through the capillary tube, and (3) a heat block of three temperature conditions A method of performing gene amplification by the PCR method by combining a cylindrical shape, winding a capillary around three cylindrical heat blocks, and flowing a sample containing the gene through the capillary is shown.

For example, Patent Document 4 discloses a method in which a temperature cycle of a PCR method is performed by providing a micro inspection chip with three temperature regions and liquid continuously traversing the temperature regions.
JP 2004-28589 A JP 2006-149379 A Japanese Patent No. 3120466 Special table 2004-532003 gazette

  However, in the method of Patent Document 3, the method using three temperature vessels requires a large space for the temperature vessel, and the inspection apparatus becomes large, and the method using a heat block is long because the temperature cycle is repeated. Capillary tubes are necessary, and the inspection apparatus is also enlarged.

  Also, in the method of Patent Document 4, in order to repeat the temperature cycle, a long flow path is required on the micro test chip, and the micro test chip becomes large.

  This invention is made | formed in view of the said situation, and it aims at providing the test | inspection apparatus which can perform gene amplification by PCR method using a small micro test | inspection chip.

  The object of the present invention can be achieved by the following constitution.

1. A micro inspection chip,
In an inspection apparatus that is connected to the micro inspection chip and includes a liquid supply unit for supplying a liquid inside the micro inspection chip,
A temperature variable section having a plurality of temperature regions;
An inspection apparatus comprising: a moving portion that relatively moves the temperature variable portion and the micro inspection chip.

  2. 2. The inspection apparatus according to 1, further comprising a separation connecting portion that detachably connects the liquid feeding portion and the micro inspection chip.

  3. 3. The inspection apparatus according to 2, wherein the liquid feeding unit and the micro test chip are separated by relatively moving the liquid feeding unit in a direction perpendicular to the surface of the micro test chip.

  4). 4. The moving unit according to any one of claims 1 to 3, wherein the moving unit moves the temperature variable unit and the micro test chip in a direction parallel to the surface of the micro test chip and relatively moves them. Inspection equipment.

  5). 4. The moving unit according to any one of 1 to 3, wherein the moving unit rotates and relatively moves the temperature variable unit and the micro test chip about an axis perpendicular to the surface of the micro test chip. The inspection device described.

6). A detection unit for detecting a reaction result inside the micro inspection chip;
6. The moving unit according to claim 1, wherein the moving unit moves the detecting unit relative to the micro test chip in conjunction with a relative movement between the temperature variable unit and the micro test chip. The inspection device described in 1.

  7. 7. The inspection apparatus according to any one of 1 to 6, wherein the liquid feeding unit is a micro pump.

  According to the present invention, the micro inspection chip is provided with a liquid supply section for supplying liquid in the micro inspection chip, and the temperature variable section having a plurality of temperature regions and the micro inspection chip are moved relative to each other. The temperature cycle can be repeated with the liquid stopped. Accordingly, it is possible to provide a test apparatus that can perform gene amplification by the PCR method using a small micro test chip without requiring a long channel on the micro test chip.

  Hereinafter, the present invention will be described based on the illustrated embodiment, but the present invention is not limited to the embodiment. In the drawings, the same or equivalent parts are denoted by the same reference numerals, and redundant description is omitted.

  First, the inspection apparatus of the present invention will be described with reference to FIGS. 1 and 2. FIG. 1 is a perspective view showing an example of an inspection apparatus 1.

  In FIG. 1, the inspection apparatus 1 includes a tray 3, a transport port 5, an operation unit 7, a display unit 9, and the like. A micro test chip 100 into which a specimen, a reagent, and the like have been injected in advance is set on the tray 3, and is transported from the transport port 5 into the test apparatus 1 by a loading mechanism (not shown) for testing. Inspection contents, inspection target data, and the like are input to the inspection apparatus 1 using the operation unit 7, and inspection results are displayed on the display unit 9.

  FIG. 2 is a block diagram showing an example of the internal configuration of the inspection apparatus 1 shown in FIG.

  In FIG. 2, the inspection apparatus 1 includes a liquid feeding unit 210, a temperature variable unit 230, a detection unit 250, a drive control unit 270, a moving unit 280, a separation connection unit 290, and the like. The micro inspection chip 100 is transported into the inspection apparatus 1 from the transport port 5 by a loading mechanism (not shown), and is connected to the liquid feeding section 210 by the separation connection section 290. By the liquid feeding by the liquid feeding unit 210, the specimen and the reagent in the micro test chip 100 are mixed and reacted to perform tests such as target substance detection and disease determination.

  The micro inspection chip 100 is equivalent to what is generally called an analysis chip, a microreactor chip, and the like. For example, the micro inspection chip 100 is made of resin, glass, silicon, ceramics, and the like. Is formed with a fine flow path having a level of several μm to several hundred μm. The size and shape of the micro inspection chip 100 is usually a plate shape having a length and width of several tens of mm and a thickness of several mm. An example of the configuration of the micro inspection chip 100 will be described in detail with reference to FIG.

  The liquid feeding unit 210 is a unit for feeding liquid in the micro test chip 100, and includes a micro pump 211, a chip connection unit 213, a driving liquid tank 215, a driving liquid supply unit 217, and the like. The liquid feeding unit 210 includes one or a plurality of micropumps 211. The micropump 211 feeds the liquid in the micro test chip 100 by injecting or sucking the driving liquid 216 into the micro test chip 100. The chip connection unit 213 connects the micropump 211 and the micro inspection chip 100.

  The driving liquid supply unit 217 supplies the driving liquid 216 from the driving liquid tank 215 to the micropump 211. The driving liquid tank 215 can be removed and replaced from the driving liquid supply unit 217 to replenish the driving liquid 216. One or a plurality of pumps are formed on the micropump 211, and the plurality of pumps can be driven independently or in conjunction with each other.

  The micro inspection chip 100 and the micropump 211 are connected by a chip connection part 213 and communicated with each other. When the micro pump 211 is driven, the driving liquid 216 is injected or sucked into the micro test chip 100 from the micro pump 211 via the chip connection portion 213. Samples, reagents, and the like housed in a plurality of housing parts in the micro test chip 100 are sent in the micro test chip 100 by the driving liquid 216.

  The temperature variable unit 230 has a plurality of temperature regions (for example, three temperature regions of a heat denaturation temperature of about 95 ° C., an annealing temperature of about 55 ° C., and a polymerization temperature of about 70 ° C.). Do. Each temperature region is configured by an element or the like whose temperature can be variably controlled by energization of a heater, a Peltier element, or the like.

  In the following description, the temperature variable unit 230 will be described as having three temperature regions, but the temperature region is not limited to three. For example, in the case of a method of performing gene amplification in two temperature regions (for example, 90 ° C. and 65 ° C.) as in the shuttle PCR method, the temperature variable unit 230 may have two temperature regions. What is necessary is just to consider that it has the temperature range optimal for PCR method.

  The detection unit 250 includes a light source such as a light emitting diode (LED) or a laser and a light receiving unit such as a photodiode (PD), and the like, and a target substance contained in a product liquid obtained by a reaction in the micro inspection chip 100, Optical detection is performed at the position of the detection region on the micro inspection chip 100. The arrangement of the light source and the light receiving unit includes a transmission type and a reflection type, and may be determined as necessary.

  The drive control unit 270 includes a microcomputer, a memory, and the like (not shown), and drives, controls, and detects each unit in the inspection apparatus 1.

  The moving unit 280 makes the temperature variable unit 230 and the micro test chip 100 contact or approach each other, and moves the temperature variable unit 230 and the micro test chip 100 relative to each other, thereby changing the temperature range of the temperature variable unit 230. The temperature of the micro test chip 100 is changed, and gene amplification is performed by the PCR method. The moving unit 280 will be described in detail with reference to FIGS.

  The separation connecting unit 290 separates or connects the liquid feeding unit 210 and the micro test chip 100 by moving part or all of the liquid feeding unit 210 or the micro test chip 100. The separation connecting portion 290 will be described in detail with reference to FIGS. 4, 6 and 7.

  Next, an example of the configuration of the micro inspection chip 100 will be described with reference to FIG. FIG. 3 is a schematic diagram illustrating an example of the configuration of the micro test chip 100, FIG. 3A is a side view, and FIG. 3B is a top view illustrating an example of the configuration of the flow path on the micro test chip 100. FIG. 3C is a top view illustrating another example of the configuration of the flow path on the micro test chip 100.

  In FIG. 3A, a micro test chip 100 is formed of a water-repellent resin material such as polypropylene, for example, and has a groove-like flow path 110 for storing and feeding a liquid such as a reagent or a specimen. And a top plate 103 that is bonded to the surface of the flow path substrate 101 where the flow path 110 is formed and functions as a lid of the flow path 110 of the flow path substrate 101. The flow path substrate 101 and the top plate 103 are provided with a connection port 113 between the micro pump unit 210 and the micro test chip 100, an injection port 123 for injecting a specimen, an air vent hole 119, and the like. Sealed with a stop member 105.

  The channel 110 communicates with the driving liquid injection unit 111 and the injection port 123 communicated with the connection port 113, and mixes the sample storage unit 121 that stores the sample X, the plurality of reagent storage units 115, and the reagent and the sample X. A plurality of amplifying units 117 for amplifying the reaction, a liquid feeding channel 107 connecting the plurality of reagent storing units 115 and the plurality of amplifying units 117, and a sample X in the sample storing unit 121 are sent to the plurality of amplifying units 117 and divided. The sample dividing path 125 and the like are configured.

  The plurality of amplifying units 117 are changed in temperature during reaction by a temperature variable unit 230 (not shown), and the amplification result is detected by the detecting unit 250 (not shown), so that the surface of the channel substrate 101 on which the channel 110 is formed. It is formed as a deep channel reaching near the back surface. Further, at least a part of the plurality of amplifying units 117 is formed to be optically transparent for optical detection by the detection unit 250.

  FIG. 3B shows an example in which the sample X is amplified by reacting with the four types of reagents A to D. Reagents A, B, C, and D are stored in advance in each of the four reagent storage units 115. The sample X is injected and stored in the sample storage unit 121 from the injection port 123 using, for example, a pipette. After injection of the specimen X, the injection port 123 is sealed with a sheet-like sealing member 105, for example.

  Each of the four reagent storage units 115 and the sample storage unit 121 is in communication with five driving liquid injection units 111. Each of the four reagent reservoirs 115 communicates with the four amplifying units 117 via the liquid supply channel 107. The sample storage unit 121 communicates with the four liquid supply channels 107 via the sample dividing channel 125.

  The driving liquid 216 is injected into the driving liquid injection part 111 from the micropump 211 connected to the four reagent storage parts 115 and the specimen storage part 121 via the connection port 113 and the driving liquid injection part 111. As a result, the specimen X is divided into four through the specimen dividing path 125 and fed to each of the four liquid feeding passages 107. At the same time, the reagents A, B, C, and D stored in each of the four reagent storage units 115 are also supplied to each of the four liquid supply channels 107. As a result, the sample X divided into four and the four reagents A, B, C, and D are injected into each of the four amplifying units 117 while being mixed in the four liquid supply channels 107. The air in the four amplifying units 117 is discharged to the outside from the air vent hole 119.

  In this way, the sample X and the reagents A to D are mixed, and the amplification A to D of the sample X and the reagents A to D are performed in each of the four amplification units 117. At this time, the temperature of the mixed solution of the specimen X and the reagents A to D is changed by the temperature variable unit 230 (not shown), and amplification is promoted. After the amplification is completed, the reaction result between the sample X and the reagents A to D is detected by the detection unit 250 (not shown). In addition, although the example which uses the amplification part 117 also for the detection by the detection part 250 was shown in this example, you may provide the flow path (reaction detection part) for a detection separately. At this time, after the amplification is completed, the mixed solution of the specimen X and the reagents A to D is sent from the amplification unit 117 to the reaction detection unit, and then detection is performed.

  FIG. 3C shows an example in which the amplification unit 117 is also used for mixing the sample X and the four types of reagents A to D. Similarly to the example of FIG. 3B, the reagents A, B, C, and D are stored in advance in each of the four reagent storage units 115. The sample X is injected and stored in the sample storage unit 121 from the injection port 123 using, for example, a pipette. After injection of the specimen X, the injection port 123 is sealed with a sheet-like sealing member 105, for example.

  Each of the four reagent storage units 115 and the sample storage unit 121 is in communication with five driving liquid injection units 111. Each of the four reagent reservoirs 115 communicates with the four amplifying units 117 via the liquid supply channel 107. The sample storage unit 121 communicates with the four amplification units 117 via the sample dividing path 125.

  First, the driving liquid 216 is injected into the driving liquid injection unit 111 from the micropump 211 connected to the sample storage unit 121 via the connection port 113 and the driving liquid injection unit 111, so that the sample X is divided into the samples. It is divided into four through the path 125 and injected into each of the four amplifying units 117. The air in the four amplifying units 117 is discharged to the outside from the air vent hole 119.

  Next, the driving liquid 216 is injected into the four driving liquid injection units 111 from the micropump 211 connected to each of the four reagent storage units 115 via the connection port 113 and the driving liquid injection unit 111. Reagents A to D are injected into each of the four amplifying units 117 through the liquid supply channel 107. The air in the four amplifying units 117 is discharged to the outside from the air vent hole 119.

  In this way, the sample X and the reagents A to D are mixed, and the amplification A to D of the sample X and the reagents A to D are performed in each of the four amplification units 117. At this time, the temperature of the mixed solution of the specimen X and the reagents A to D is changed by the temperature variable unit 230 (not shown), and amplification is promoted. After the amplification is completed, the reaction result between the sample X and the reagents A to D is detected by the detection unit 250 (not shown).

(First embodiment)
Next, a first embodiment of the temperature variable unit 230, the moving unit 280, and the separation connecting unit 290 according to the present invention will be described with reference to FIGS. FIG. 4 is a schematic diagram showing the first embodiment of the temperature variable unit 230, the moving unit 280, and the separation connecting unit 290. FIG. 4A shows a state in which the micro inspection chip 100 is transported into the inspection apparatus 1. FIG. 4B shows a state in which the temperature of the amplifying unit 117 is changed.

  In FIG. 4A, when the micro inspection chip 100 is transported into the inspection apparatus 1 by a loading mechanism (not shown), the micro inspection chip 100 is lifted in the direction of the arrow A in the drawing by the separation connection portion 290, and the chip connection portion 213. And the connection port 113 of the micro inspection chip 100 are connected. A packing 213 a is provided at a portion of the chip connection portion 213 that contacts the connection port 113, thereby preventing leakage from the contact portion between the chip connection portion 213 and the connection port 113.

  Next, the driving liquid 216 is injected into the micro test chip 100 from the micro pump 211 via the chip connection part 213 and the connection port 113. When the specimen X and the reagents A to D are fed by the driving liquid 216 and mixed by the amplification unit 117, the micropump 211 is stopped.

  The moving unit 280 has a conveyor unit 280a having a belt conveyor shape, and the temperature variable unit 230 is mounted on the transport unit 280a. The temperature variable unit 230 has a plurality of temperature regions necessary for the PCR method, for example. Here, the temperature variable unit 230 has, for example, three temperature regions, where the region 230a has a heat denaturation temperature of about 95 ° C., the region 230b has an annealing temperature of about 55 ° C., and the region 230c has a polymerization temperature of about 70 ° C. Initially, the temperature variable unit 230 is not energized, and the three temperature regions are not set to predetermined temperatures.

  In FIG. 4B, the temperature variable unit 230 is energized, and each of the three temperature regions is controlled to a predetermined temperature. With the three temperature regions reaching a predetermined temperature, the transport unit 280a of the moving unit 280 is lifted in the direction of arrow B in the figure, and the region 230a of the temperature variable unit 230 flows to the amplifying unit 117 of the micro test chip 100. It is brought into contact with the back surface side of the road forming surface or with a slight gap. As a result, the liquid mixture of the sample and the reagent in the amplification unit 117 is held at the temperature of the region 230a, and the reaction is promoted.

  After a predetermined time has elapsed, the moving unit 280 is rotated and moved in the direction of arrow C in the figure, and the region 230b of the temperature variable unit 230 is in contact with the back surface side of the flow path forming surface of the flow path substrate 101 of the micro test chip 100 or slightly The mixture is approached with a gap, and the liquid mixture of the specimen and the reagent in the amplification unit 117 is held at the temperature of the region 230b, and the reaction is promoted.

  Similarly, after elapse of a predetermined time, the region 230c is brought into contact with or close to the back surface side of the flow path forming surface of the flow path substrate 101 of the micro test chip 100, and the sample and the reagent in the amplification unit 117 are approached. Is maintained at the temperature of the region 230c, and the reaction is promoted. This is repeated until the amplification by the PCR method is completed.

  FIG. 5 is a schematic diagram showing a modification of the first embodiment described above. FIG. 5A shows an example in which the moving unit 280 is continuously driven, and FIG. The example which provided the detection part 250 is shown.

  In FIG. 5A, each of the three regions 230a, 230b, and 230c provided on the transport unit 280a has a length proportional to the time during which the amplification unit 117 should be held at each temperature by the PCR method. Yes. Accordingly, the temperature cycle necessary for the PCR method can be realized by continuously rotating the transport unit 280a in the direction of arrow C in the figure at a constant speed.

  In FIG. 5B, a detection unit 250 is provided on the transport unit 280a in addition to the three regions 230a, 230b, and 230c. The detection unit 250 is, for example, a reflection type detection unit, and a light source and a light receiving unit are provided on the same substrate. By adopting such a configuration, the amplification unit 117 is maintained at the temperature of the three regions 230a, 230b, and 230c as in the first embodiment, and the reaction is promoted by the detection unit 250 after the reaction is promoted. Amplification can be performed while monitoring the progress of the reaction in real time, such as monitoring and further promoting the reaction.

  As described above, according to the first embodiment, the temperature variable unit provided on the moving unit is in contact with the amplifying unit of the micro test chip or in a state of being close to the amplifier with a slight gap, By moving intermittently or continuously with respect to the amplification unit, the temperature cycle can be repeated in a state where the mixed solution of the specimen and the reagent is stopped in the amplification unit. Accordingly, it is possible to provide a test apparatus that can perform gene amplification by the PCR method using a small micro test chip without requiring a long channel on the micro test chip.

(Second Embodiment)
Next, a second embodiment of the temperature variable unit 230, the moving unit 280, and the separation connection unit 290 according to the present invention will be described with reference to FIG. FIG. 6 is a schematic diagram showing a second embodiment of the temperature variable unit 230, the moving unit 280, and the separation connecting unit 290. FIG. 6A is a top view of the micro test chip 100 as viewed from the top plate 103 side. FIG. 6B shows a side view.

  6 (a) and 6 (b), the second embodiment is different from the first embodiment in that the transport unit 280a of the moving unit 280 has a disk shape instead of a belt conveyor shape. It is a point. Other points are the same as those in the first embodiment. The transport unit 280a can rotate in the direction of arrow D around the rotation shaft 280b, and the temperature variable unit 230 is mounted on the transport unit 280a.

  The temperature variable unit 230 has a plurality of temperature regions necessary for the PCR method. Here, as in the first embodiment, the temperature variable unit 230 has, for example, three temperature regions, the region 230a has a heat denaturation temperature of about 95 ° C., the region 230b has an annealing temperature of about 55 ° C., and the region 230c has The polymerization temperature is about 70 ° C.

  In this example, similarly to the example shown in FIG. 5A, the three regions 230a, 230b, and 230c are held at each temperature by the amplification unit 117 by the PCR method in the rotational movement direction around the rotation shaft 280b. It has a length proportional to the time to be done. Accordingly, the temperature and time cycle required for the PCR method can be realized by continuously rotating the transport unit 280a in the direction of arrow D in the figure at a constant speed.

  Of course, the three regions 230a, 230b, and 230c may have the same configuration as the example shown in FIGS. 4 and 5B.

  Next, the operation of the second embodiment will be described. In FIG. 6B, when the micro inspection chip 100 is conveyed into the inspection apparatus 1 by a loading mechanism (not shown), the micro inspection chip 100 is lifted in the direction of the arrow A in the drawing by the separation connection portion 290, and the chip connection portion 213. And the connection port 113 of the micro inspection chip 100 are connected. A packing 213 a is provided at a portion of the chip connection portion 213 that contacts the connection port 113, thereby preventing leakage from the contact portion between the chip connection portion 213 and the connection port 113.

  Next, the driving liquid 216 is injected into the micro test chip 100 from the micro pump 211 via the chip connection part 213 and the connection port 113. When the specimen X and the reagents A to D are fed by the driving liquid 216 and mixed by the amplification unit 117, the micropump 211 is stopped.

  As described above, the moving unit 280 has the disk-shaped transport unit 280a, and the temperature variable unit 230 is mounted on the transport unit 280a. Initially, the temperature variable unit 230 is not energized, and the three temperature regions are not set to predetermined temperatures.

  Subsequently, the temperature variable unit 230 is energized, and each of the three temperature regions is controlled to a predetermined temperature. With the three temperature regions reaching the predetermined temperatures, the moving unit 280 is lifted in the direction of arrow B in the figure, and the region 230a of the temperature variable unit 230 is formed on the flow path forming surface of the amplification unit 117 of the micro test chip 100. It approaches the back side with a slight gap. A cycle of temperature and time required for the PCR method can be realized by continuously rotating the transport unit 280a in the direction of arrow D in the figure at a constant speed. This is repeated until the amplification by the PCR method is completed.

  As described above, according to the second embodiment, the temperature variable portion provided on the disk-shaped moving portion is brought into contact with the amplifying portion of the micro test chip or with a slight gap therebetween. In this state, the temperature cycle can be repeated in a state where the liquid mixture of the specimen and the reagent is stopped in the amplification unit by moving the amplification unit intermittently or continuously. Accordingly, it is possible to provide a test apparatus that can perform gene amplification by the PCR method using a small micro test chip without requiring a long channel on the micro test chip.

(Third embodiment)
Next, a third embodiment of the temperature variable unit 230, the moving unit 280, and the separation connecting unit 290 according to the present invention will be described with reference to FIG. FIG. 7 is a schematic diagram illustrating a third embodiment of the temperature variable unit 230, the moving unit 280, and the separation connection unit 290. FIG. 7A is a top view of the micro inspection chip 100 as viewed from the top plate 103 side. FIG. 7B is a cross-sectional view taken along the line EE ′ of FIG.

  7A and 7B, the third embodiment is different from the first and second embodiments in that the micro inspection chip 100 is rotated instead of moving the temperature variable unit 230. It is a point to move.

  First, the operation of the third embodiment will be described. In FIG. 7B, in the initial state, the chip connection portion 213 is pulled up in the direction opposite to the arrow A in the figure by the separation connection portion 290 and separated from the micropump 211.

  When the micro inspection chip 100 is conveyed into the inspection apparatus 1 by a loading mechanism (not shown), the chip connection portion 213 is pushed down in the direction of arrow A in the figure by the separation connection portion 290. Thereby, the chip connection part 213 and the connection port 113 of the micro test chip 100, and the chip connection part 213 and the micropump 211 are connected (state of broken line in FIG. 7B).

  Packing 213 a is provided in the portion of the chip connection portion 213 that contacts the connection port 113 and the portion that contacts the micropump 211, and the contact portion between the chip connection portion 213 and the connection port 113 and the chip connection portion 213 and the micropump 211. Prevents leakage from the contact area.

  Next, the driving liquid 216 is injected into the micro test chip 100 from the micro pump 211 via the chip connection part 213 and the connection port 113. When the specimen X and the reagents A to D are fed by the driving liquid 216 and mixed by the amplification unit 117, the micropump 211 is stopped. Subsequently, the chip connection part 213 is pulled up in the direction opposite to the arrow A in the figure by the separation connection part 290, and is separated from the micropump 211 and the micro test chip 100 (solid line state in FIG. 7B).

  The holding unit 300 has a disk shape, and the temperature variable unit 230 and the detection unit 250 are mounted thereon. In the vicinity of the center of the holding unit 300, a moving unit 280 for rotating the micro test chip 100 relative to the holding unit 300 is provided. Initially, the temperature variable unit 230 is not energized, and the three temperature regions are not set to predetermined temperatures.

  Subsequently, the temperature variable unit 230 is energized, and each of the three temperature regions is controlled to a predetermined temperature. The moving unit 280 and the holding unit 300 are lifted in the direction of arrow B in the drawing in a state where the three temperature regions have reached predetermined temperatures, respectively. As a result, the moving unit 280 is inserted into the notch 101 a provided in the flow path substrate 101 of the micro test chip 100, and the micro test chip 100 can be rotated by the moving unit 280.

  In addition, the region 230a of the temperature variable unit 230 is brought into contact with the back surface side of the flow path forming surface of the amplifying unit 117 of the micro test chip 100 or is approached with a slight gap. As a result, the liquid mixture of the sample and the reagent in the amplification unit 117 is held at the temperature of the region 230a, and the reaction is promoted.

  Next, after a predetermined time has elapsed, the micro inspection chip 100 is rotated 90 ° in the direction of arrow F in FIG. 7A by the moving unit 280, and the back surface of the flow path forming surface of the amplifying unit 117 is the region of the temperature variable unit 230. 230b is abutted or approached with a slight gap. As a result, the liquid mixture of the sample and the reagent in the amplification unit 117 is held at the temperature of the region 230b, and the reaction is promoted. Similarly, after a predetermined time elapses, the micro inspection chip 100 is rotated 90 ° in the direction of arrow F in the figure by the moving unit 280, or approached with a slight gap. As a result, the liquid mixture of the sample and the reagent in the amplification unit 117 is held at the temperature of the region 230c, and the reaction is promoted.

  Next, the micro inspection chip 100 is rotated 90 ° in the direction of arrow F in the figure by the moving unit 280, and the back surface of the flow path forming surface of the amplifying unit 117 is brought into contact with the detecting unit 250 or approached with a slight gap. The The detection unit 250 is, for example, a reflection type detection unit, and a light source and a light receiving unit are provided on the same substrate. With this configuration, after the amplification unit 117 is maintained at the temperature of the three regions 230a, 230b, and 230c and the reaction is promoted, the detection unit 250 monitors the progress of the reaction and further promotes the reaction. Thus, amplification can be performed while monitoring the progress of the reaction in real time. This is repeated until the amplification by the PCR method is completed.

  Here, the chip connection unit 213 is separated from the micropump 211, and has been described as being moved up and down in the direction of arrow A in the figure by the separation connection unit 290. However, the present invention is not limited to this. The pump 211 may be integrated, and may be moved up and down in the direction of arrow A in the figure together by the separation connecting portion 290.

  As described above, according to the third embodiment, after the liquid mixture of the specimen and the reagent is fed to the amplifying unit, the micro test chip is kept at a temperature in a state where the micro test chip and the micro pump are separated. By rotating and moving with respect to the variable part, the temperature cycle can be repeated in a state where the mixed liquid of the specimen and the reagent is stopped in the amplification part. Accordingly, it is possible to provide a test apparatus that can perform gene amplification by the PCR method using a small micro test chip without requiring a long channel on the micro test chip.

  As described above, according to the present invention, the micro inspection chip is provided with a liquid supply section for supplying a liquid in the micro inspection chip, and the temperature variable section having a plurality of temperature regions and the micro inspection chip are relatively moved. Thus, the temperature cycle can be repeated with the liquid stopped in the micro test chip. Accordingly, it is possible to provide a test apparatus that can perform gene amplification by the PCR method using a small micro test chip without requiring a long channel on the micro test chip.

  It should be noted that the detailed configuration and detailed operation of each component constituting the inspection apparatus according to the present invention can be changed as appropriate without departing from the spirit of the present invention.

It is a perspective view which shows an example of an inspection apparatus. It is a block diagram which shows an example of an internal structure of a test | inspection apparatus. It is a schematic diagram which shows an example of a structure of a micro test | inspection chip. It is a schematic diagram which shows 1st Embodiment of a temperature variable part, a moving part, and the isolation | separation connection part. It is a schematic diagram which shows the modification of 1st Embodiment. It is a schematic diagram which shows 2nd Embodiment of a temperature variable part, a moving part, and the isolation | separation connection part. It is a schematic diagram which shows 3rd Embodiment of a temperature variable part, a moving part, and the isolation | separation connection part.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Inspection apparatus 3 Tray 5 Conveyance port 7 Operation part 9 Display part 100 Micro test | inspection chip 101 Flow path board | substrate 103 Top plate 105 Sealing member 107 Liquid supply flow path 110 Flow path 111 Drive liquid injection | pouring part 113 Connection port 115 Reagent storage part 117 Amplifying section 119 Air vent hole 121 Sample storage section 123 Inlet 125 Sample dividing path 210 Liquid feeding section 211 Micro pump 213 Chip connection section 213a Packing 215 Driving liquid tank 216 Driving liquid 217 Driving liquid supply section 230 Temperature variable section 230a area 230b area 230c Area 250 Detecting unit 270 Drive control unit 280 Moving unit 280a Conveying unit 280b Rotating shaft 290 Separation connecting unit 300 Holding unit

Claims (7)

A micro inspection chip,
In an inspection apparatus that is connected to the micro inspection chip and includes a liquid supply unit for supplying a liquid inside the micro inspection chip,
A temperature variable section having a plurality of temperature regions;
An inspection apparatus comprising: a moving portion that relatively moves the temperature variable portion and the micro inspection chip. The inspection apparatus according to claim 1, further comprising a separation connecting portion that detachably connects the liquid feeding portion and the micro inspection chip. The inspection apparatus according to claim 2, wherein the liquid feeding unit and the micro test chip are separated by relatively moving the liquid feeding unit in a direction perpendicular to the surface of the micro test chip. The said moving part moves the said temperature variable part and the said micro test | inspection chip | tip linearly in one direction parallel to the surface of the said micro test | inspection chip, and makes it move relatively. The inspection device described in 1. The said moving part rotates the said temperature variable part and the said micro test | inspection chip | tip around the axis | shaft perpendicular | vertical to the surface of the said micro test | inspection chip, It moves relative to any one of Claims 1 thru | or 3 characterized by the above-mentioned. Inspection device according to item. A detection unit for detecting a reaction result inside the micro inspection chip;
6. The moving unit according to claim 1, wherein the moving unit moves the detecting unit relative to the micro test chip in conjunction with a relative movement between the temperature variable unit and the micro test chip. The inspection apparatus according to item 1. The inspection apparatus according to claim 1, wherein the liquid feeding unit is a micropump.

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