JP2017192341A - Pre-treatment kit for gene analysis, nucleic acid analysis chip, and gene analysis system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a chip for nucleic acid analysis that ensures ease of handling that can be attained even by a user without expertise in medical treatment and can further improve the accuracy of biochemical examination.SOLUTION: According to the present invention, there are provided a pre-treatment kit for gene analysis, a nucleic acid analysis chip, and gene analysis system, each of which comprising: a PCR channel 34 into which a liquid containing genomic DNA extracted from a collected product is injected and amplifies a nucleic acid fragment of a target nucleic acid, while flowing it downstream; a wash channel 32 through which at least a cleaning wash flows; a hybridization channel 37 communicated with the PCR channel 34 and the wash channel 32, the hybridization channel causing hybridization of an amplified product flowing from the PCR cannel 34 and flowing the amplified product downward; and a chip-measuring device 4 for identifying a target nucleic acid by measuring a hybridization signal generated in the hybridization channel 37. After the hybridization, the hybridization channel 37 can be cleaned by flowing a washing solution from the wash channel 32.SELECTED DRAWING: Figure 5

Description

  The present invention relates to a pretreatment kit for gene analysis for extracting a biological material from a sample collected from any one of the human body, excrement, and soil, and for nucleic acid analysis for analyzing a target nucleic acid to be detected from such sample. The present invention relates to a chip and a gene analysis system.

  Conventionally, medical examinations have been carried out exclusively at specialized medical institutions such as hospitals, but in Japan, with the arrival of a super-aging society, the burden of medical examinations at medical institutions has become serious. For this reason, tests that require dedicated devices such as X-ray imaging and electrocardiogram measurements, and tests that require a syringe, such as blood tests, will continue to be performed at medical institutions, while users can easily perform tests. It is necessary to consider reducing the burden of screening at medical institutions by using self-examination at home. If such self-examination performed at home can be established, therapeutic medical care and preventive medical care can be enhanced even in developing countries where necessary medical services are not sufficiently delivered.

  Under such circumstances, in order to enable next-generation self-examination, in addition to downsizing, generalization, and cost reduction of the inspection device, it can be realized even by users who do not have specialized knowledge about medical care. A certain level of ease of handling is required.

  In response to such social needs, in recent years, gene analysis using a nucleic acid analysis chip (DNA chip) has been realized. Nucleic acid analysis chips are used in various fields such as the medical field, chemical field, drug discovery field, clinical laboratory field, food field, agricultural field, engineering field, forensic field, and criminal field, such as the medical field. In general, screening of various diseases such as cancer is performed by using a biochemical sample such as blood.

  A nucleic acid analysis chip is a device in which nucleic acid probes are immobilized at high density on a glass or silicon substrate. When gene analysis is performed with high accuracy using a nucleic acid analysis chip, gene amplification is required in advance. If the nucleic acid amplified in this way has a complementary base sequence with the nucleic acid probe on the substrate in the next hybridization step, it binds to it. Next, in the washing step, the nucleic acid analysis chip after the hybridization reaction is washed with a predetermined washing solution. As a result, all DNA samples not bound to the nucleic acid probe on the substrate are washed away.

  Subsequently, the presence and / or amount of the target nucleic acid is detected by scanning the hybridization signal from the washed nucleic acid analysis chip. In this scanning, a laser beam having a wavelength suitable for exciting the fluorescent material is irradiated onto the substrate of the nucleic acid analysis chip and scanned. As a result, the amount of luminescence of the nucleic acid bound to each spotted nucleic acid probe is measured, and based on this, analysis processing can be performed.

  In the nucleic acid analysis chip as described above, for example, a technique disclosed in Patent Document 1 is disclosed for the purpose of meeting the needs for self-examination performed at home. This Patent Document 1 discloses a nucleic acid analysis chip that can detect a reaction instantaneously, has a small and portable analyzer configuration, stores a reaction reagent in advance, and can be disposed of together with the reagent after analysis. Patent Document 2 also proposes a disposable paper-based microfluidic device for detecting and analyzing a sample in blood or urine collected from a subject.

JP 2004-28589 A Special table 2014-529083 gazette

  However, the above-described conventional nucleic acid analysis chip has room for further improvement from the viewpoint of improving the accuracy of biochemical tests. In addition, when performing an enzyme reaction or biochemical reaction with a nucleic acid analysis chip fixed to the analyzer, it may be difficult to control the temperature accurately on the nucleic acid analysis chip, and the target reaction may not proceed sufficiently. there were.

  Patent Document 1 proposes that a nucleic acid analysis chip is provided with an adsorption groove, and the chip is adsorbed and fixed at a desired position by vacuum suction through the adsorption groove by a vacuum exhaust pump. However, when such a configuration is adopted, it becomes a large barrier in reducing the size of the analyzer.

  Accordingly, the present invention has been devised in view of the above-described problems, and the object of the present invention is to reduce the size and versatility of a testing device in a nucleic acid analysis chip that enables next-generation self-examination. In addition to lowering the price, a chip for nucleic acid analysis and gene analysis that can ensure the ease of handling that can be realized even by users who have no specialized knowledge about medical care, and can further improve the accuracy of biochemical tests It is an object of the present invention to provide a pretreatment kit for gene analysis for extracting a biological material from a sample collected from any one of a human body, excrement, and soil in a stage before performing the analysis of the system.

  The pretreatment kit for gene analysis according to the first invention is a pretreatment kit for gene analysis for extracting a biological material from a sample collected from any one of a human body, excrement, and soil, and is filled with a plurality of granules. When the rod-like body in which the liquid is immersed in the granular material and the collected material is attached to the tip is inserted, the collected material is collected by bringing the tip into contact with the granular material and A collection cartridge for impregnating with a liquid, and a discharge means for discharging the liquid sent from the collection cartridge to the outside.

  A pretreatment kit for gene analysis according to a second aspect of the present invention includes the first filter for separating at least serum from the liquid sent from the attached collection cartridge and the discharging means in the first aspect. The apparatus further includes a pretreatment main body, and the discharge means discharges the liquid that has passed through the first filter to the outside.

  The pretreatment kit for gene analysis according to the third invention is characterized in that, in the first or second invention, the liquid contains a bactericidal component, a gene detection primer, and a polymerase enzyme.

  The pretreatment kit for gene analysis according to a fourth invention is the pretreatment kit for gene analysis according to the third invention, wherein the pretreatment main body further has a second filter for separating at least viruses and bacteria from the liquid that has passed through the first filter. The discharge port discharges the liquid that has passed through the second filter to the outside.

  A pretreatment kit for gene analysis according to a fifth invention is the pretreatment kit for gene analysis according to any one of the first to fourth inventions, wherein the sampling cartridge is biased in advance in a narrowing direction and the tip of the rod-like body is It has an insertion port to be inserted, the inside of the insertion port is filled with the granular material, and the liquid is immersed therein.

  The pretreatment kit for gene analysis according to a sixth invention is the pretreatment kit for gene analysis according to the fifth invention, wherein the rod-like body has a protruding portion that protrudes outward from a base portion of a tip to which the collected material is attached. It is locked to the insertion port.

  A pretreatment kit for gene analysis according to a seventh aspect of the present invention is the pretreatment kit according to any one of the first to sixth aspects, wherein the granular material is filled when the collection cartridge and the pretreatment main body are mounted. The container and the first filter are separated from each other through a laminated body in which a network body through which the liquid can pass and a film through which the liquid cannot pass are laminated, and when the liquid is delivered from the sampling cartridge, The film is formed with holes for allowing the liquid to pass therethrough, and the liquid is allowed to pass through, and the granular material is held by the mesh.

  In the pretreatment kit for gene analysis according to the eighth invention, in any one of the first invention to the seventh invention, the granular material is composed of any one of metal, ceramics, and resin, and on the surface thereof, Any one of zeolite, emulsion gel, hydrophilic polymer, and amino acid substance is formed.

  The pretreatment kit for gene analysis according to the ninth invention is a pretreatment kit for gene analysis for extracting a biological material from a sample collected from any one of a human body, excrement and soil, wherein the epidermis is composed of an elastic material. In addition, a liquid containing a bactericidal component, a gene detection primer, and a polymerase enzyme was enclosed in the epidermis, and a rod-like body with the collected material attached to the tip was inserted. In this case, the collected material is collected by bringing the tip into contact with the granular material, and the collecting cartridge is impregnated with the liquid that has flowed out by breaking the epidermis by the tip, and is sent from the collecting cartridge. And a discharging means for discharging the liquid that has been discharged to the outside.

  A nucleic acid analysis chip according to a tenth aspect of the invention is a nucleic acid analysis chip for analyzing a target nucleic acid to be detected from a sample collected from any one of a human body, excrement, and soil, a gene detection primer, a polymerase enzyme, A liquid containing a biological material extracted from the collected material is injected, and a PCR channel that flows downstream while amplifying nucleic acid fragments in the target nucleic acid, and an amplification that continues from the PCR channel and flows in from the PCR channel It comprises a hybridization channel for allowing a product to hybridize while flowing downstream, and a target nucleic acid analyzing means for identifying the target nucleic acid by measuring a hybridization signal generated in the hybridization channel. And

  The nucleic acid analysis chip according to an eleventh aspect of the present invention is the nucleic acid analysis chip according to the tenth aspect of the present invention, further comprising a wash channel through which at least a washing solution flows. The hybridization channel is also continuous from the wash channel, and after the hybridization, It is characterized in that cleaning can be performed by flowing a cleaning liquid from the wash channel.

  The nucleic acid analysis chip according to a twelfth aspect of the present invention is the tenth or eleventh aspect of the present invention, wherein the PCR channel is configured with a micron order diameter, and the heating temperature provided at predetermined intervals is 30 to 150 ° C. The target nucleic acid is disposed so as to pass over a certain heating wire, and the target nucleic acid is thermally denatured when passing through the heating wire.

  The nucleic acid analysis chip according to a thirteenth aspect of the present invention is the nucleic acid analysis chip according to the twelfth aspect of the present invention, wherein the PCR flow path is folded after passing over a heating wire provided at a predetermined interval, further passing over the heating wire, and further folded. It arrange | positions so that passing on the said heating wire may be repeated.

  The nucleic acid analysis chip according to a fourteenth aspect of the present invention is characterized in that, in the eleventh aspect, the wash flow path is alternately flowed with a cleaning solution and air.

  The nucleic acid analysis chip according to a fifteenth aspect of the invention is any one of the tenth to fourteenth aspects, further comprising a communication means for transmitting the information detected by the target nucleic acid analysis means to the outside via a communication network. Features.

  A gene analysis system according to a sixteenth invention comprises the pretreatment kit for gene analysis of any of the first to ninth inventions, and the nucleic acid analysis chip of any of the tenth to fifteenth inventions, wherein the gene The liquid discharged from the discharge port in the analysis pretreatment kit flows into the PCR flow path in the nucleic acid analysis chip.

  A biomaterial analysis chip according to a seventeenth aspect of the present invention is a biomaterial analysis chip for analyzing a biomaterial to be detected from a sample collected from any one of a human body, excrement, and soil, and extracted from the collected sample. An infusion channel in which a liquid containing a biological material is injected and flows downstream, and a hybridization that is continuous from the infusion channel and flows downstream while allowing the biological material flowing in from the infusion channel to hybridize And a target nucleic acid analyzing means for identifying the target nucleic acid by measuring a hybridization signal generated in the hybridization channel, and a wash channel for flowing at least a washing solution. It continues from the wash channel, and after the hybridization, Wherein the cleaning liquid is capable washed by being flowed.

  According to the present invention having the above-described configuration, the user collects a sample from a human body, excrement, or soil with a rod-shaped body, and the sample is attached to the tip of the sampling cartridge in which the liquid is immersed. Insert a stick-shaped body. Other than that, simply attaching the collection cartridge to the pretreatment body, and connecting the tip of the discharge port of the pretreatment body to the sample injection port of the nucleic acid analysis chip, everything is automatically Analysis will be performed.

  For this reason, even a user who has no specialized knowledge about medical care can perform genetic analysis by himself with very simple work.

  In addition, the pretreatment kit for gene analysis, the chip for nucleic acid analysis, and the chip measuring device are not complicated, so it is possible to reduce the size, and the structure suitable for self-examination at home. Moreover, since it can be provided at a low cost, it can be purchased by an individual.

  Furthermore, according to the present invention, nucleic acids can be extracted from collected samples with high efficiency, and serum, fungi, and viruses are removed in advance through a pretreatment kit for gene analysis in order to identify nucleic acids with high accuracy. In addition, these operations can be carried out very easily without the need for special skills and knowledge.

  In addition, the washing solution can be washed by flowing the washing solution from the wash channel to the hybridization channel, and the nucleic acid analysis chip can be used over and over again. This cleaning operation can be performed very easily.

  In addition, while simplifying the device configuration as described above, improving ease of handling, and improving cost performance, the accuracy of biochemical tests can be maintained at the same level as before, so the same level at medical institutions. Can be realized at home.

It is a figure which shows the system configuration | structure of the gene-analysis system to which this invention is applied. It is a figure for demonstrating the structure of the cartridge for collection | recovery. It is a figure for demonstrating the detail of the laminated body comprised by the two-layer structure of a net-like body and a film. It is a figure for demonstrating the structure of the main body for pre-processing. It is a figure for demonstrating the structure of the chip | tip for nucleic acid analysis. It is a system configuration figure of a chip measuring device. It is a figure which shows the block configuration of the control block in a chip | tip measuring apparatus. It is a figure for demonstrating the method to analyze a gene using the gene-analysis system to which this invention is applied. It is a figure which shows the state which mounted | wore the cartridge for collection to the cartridge accommodating part in the main body for pre-processing. It is a figure which shows the state which allowed the liquid to pass through the film and to flow down to the separation part side in the main body for pretreatment. It is a figure which shows the external appearance of a case accommodation type gene analysis system. It is an expanded view of a case accommodation type gene analysis system.

  Hereinafter, embodiments for carrying out a gene analysis system to which the present invention is applied will be described in detail with reference to the drawings.

  FIG. 1 shows a system configuration of a gene analysis system 1 to which the present invention is applied. The gene analysis system 1 includes a gene analysis pretreatment kit 2, a nucleic acid analysis chip (DNA chip) 3, a chip measurement device 4 to which the nucleic acid analysis chip 3 is attached, and the chip measurement device 4. Mobile terminal 5, server 7 connected to mobile terminal 5 through public communication network 6, and central control device 8 connected to public communication network 6.

  The mobile terminal 5 is, for example, a mobile phone, a smartphone, a tablet terminal, a wearable terminal, a notebook personal computer (PC), or the like, and is a device capable of communicating via the public communication network 6 based on at least a user operation. is there. The portable terminal 5 is a device that can be carried by the user at all times, but is not limited to this, and is a concept that includes all electronic devices such as a desktop PC.

  The mobile terminal 5 can download the examination application via the public communication network 6. Thus, the user can operate the mobile terminal 5 to activate the examination application, confirm his own examination record, or confirm various proposals for future health maintenance. In the following example, a case where a smartphone is applied to the mobile terminal 5 will be described as an example.

  The public communication network 6 is an Internet network or the like to which the mobile terminal 5, the server 7, and the central control device 8 are connected via a communication line. The public communication network 6 may be a so-called optical fiber communication network. The public communication network 6 is not limited to a wired communication network, and may be realized by a wireless communication network.

  A predetermined database is constructed in the server 7. Information sent via the public communication network 6 is stored in this database. Further, the server 7 transmits the accumulated information to the mobile terminal 5 via the public communication network 6 based on a request from the mobile terminal 5.

  The central control device 8 is composed of an electronic device such as a personal computer, and accesses the server 7 via the public communication network 6 and analyzes information stored therein. The central control device 8 may be operated by a specialist such as a doctor or may utilize information stored in the server 7. The knowledge obtained by analyzing information by the central control device 8 and the analysis result itself may be transmitted to the portable terminal 5 via the public communication network 6. The portable terminal 5 can display various information sent from the central control device 8 to the user via the examination application.

  The gene analysis pretreatment kit 2 is roughly classified into a plunger 21, a collection cartridge 22, and a pretreatment main body 23.

  The plunger 21 is inserted into the collection cartridge 22 and is provided to push out the liquid contained therein. The plunger 21 has an inner cylinder portion 211 and a pusher 212. The inner cylinder part 211 is extended in the longitudinal direction, and a rubber part 213 is provided at the extended end. The pusher 212 is provided on the other end side of the plunger 21 which is different from the extended end on which the rubber portion 213 is provided, and the inner cylinder portion 211 is removed from the sampling cartridge by pressing and pressing the user's finger. 22 is configured so as to be freely insertable.

  As shown in FIG. 2, the collection cartridge 22 is filled with a plurality of granular bodies 223 in a storage container 220. The granular material 223 is immersed in the liquid L. Further, the collection cartridge 22 is further provided with an inner wall 221 bulging inward and a laminate 26 provided on the bottom surface of the container 220.

  The granular material 223 may be formed of any material such as resin, ceramics, or metal. The granular material 223 may be made of die casting when made of metal, or may be made of polystyrene, for example, when made of resin. The granular material 223 may be formed by laminating these materials in a two-layer structure. For example, a resin layer may be coated on the outer shell of a metallic core made of die casting.

  The granular body 223 will be described by taking a case where it is composed of spherical beads as an example, but is not limited thereto, and may be composed of any polyhedron such as a cube or a rectangular parallelepiped. , May be configured in a random three-dimensional shape. Moreover, this granular material 223 may be comprised by the specific shape like a hook shape, for example.

  The surface of the granular material 223 may be further coated with calcium phosphate. Through this calcium phosphate layer, it is possible to make the pH of the container 220 approach neutral. Further, the surface of the granule 223 is coated with any substance of, for example, zeolite, emulsion gel, hydrophilic polymer, and amino acid substance to adsorb feces and solids as excrement to the granule 223, It is possible to make it easy to remove excess objects with a filter.

  A liquid L is injected in advance into the container 220 filled with the granular material 223 having the above-described configuration. As a component of this liquid, for example, if the collected material from which genomic DNA as a biological material is extracted is feces, it may be composed of the composition shown in Table 1 below. If RNA is to be extracted, the liquid may be composed of the composition shown in Table 2. The capacity of each component composition is expressed in terms of% by weight relative to the total weight of the liquid L.

  Here, the lysis buffer is a stabilizing substance for preventing stool oxidation, and is made of HCl, EDTA, NaCl, SDS, or the like. The DNase buffer is a solution for preventing degradation and inactivation of genomic DNA, and is composed of DNase A (deoxyribonuclease), sodium acetate containing EDTA, and the like for inhibiting DNA degradation. DNase / RNase-treated water is water from which DNase or the like, which is a DNA degrading enzyme, has been removed, and is composed of water from which a DNA degrading enzyme and an RNA degrading enzyme have been removed.

  If the collected sample is blood or urine, it may be composed of the composition shown in Table 3 below.

  If RNA is to be extracted, the liquid may be composed of the composition shown in Table 4.

  Here, the adjustment buffer is a stabilizing substance for preventing blood coagulation, and is composed of EDTA, SDS, or the like. The buffer solution is a solution for preventing the degradation and inactivation of genomic DNA, and is constituted by mixing citric acid and disodium hydrogen phosphate. DNase / RNase-treated water is water from which DNase, which is a DNA degrading enzyme, is removed, and is composed of DNA degrading enzyme and water from which RNA degrading enzyme has been removed.

  If the collected sample is saliva, it may be composed of the composition shown in Table 5 below.

  If RNA is to be extracted, the liquid may be composed of the composition shown in Table 6.

  Here, the buffer solution is a solution for preventing the degradation and inactivation of genomic DNA, and is configured by mixing citric acid and disodium hydrogen phosphate. DNase / RNase-treated water is water from which DNase, which is a DNA degrading enzyme, is removed, and is composed of DNA degrading enzyme and water from which RNA degrading enzyme has been removed. The pronase solution is a solution for removing a protein called mucin contained in saliva, and is composed of lactose, hydroxypropylcellulose, pronase and the like.

  The inner wall 221 is composed of an elastic body such as rubber, and is configured to be elastically expandable and contractible in the radial direction of the storage container 220. The inner wall 221 has an inner diameter that gradually decreases from the upper end of the container 220 toward the lower side, and the inner diameter is the smallest in the middle stage. In the middle stage where the inner diameter is the smallest, there is provided an insertion port 229 surrounded by a protruding portion 222 further protruding inward. The inner diameter is formed so as to gradually expand from the insertion port 229 downward. As a result, the container 220 is divided into two chambers, an upper chamber 220a and a lower chamber 220b, with the insertion port 229 as a boundary via the inner wall 221.

  The above-described inner cylinder portion 211 of the plunger 21 can be inserted into the upper chamber 220a. The lower chamber 220b is filled with the granular material 223, and further the liquid L is injected. The granular material 223 and the liquid L are basically charged in the lower chamber 220b, but a part thereof may overflow into the upper chamber 220a.

  The insertion port 229 formed by the inner wall 221 may be urged in advance in a direction in which the inner diameter is narrowed. That is, the insertion port 229 may be biased in advance in the direction of the arrow in FIG.

  Incidentally, the configuration of the inner wall 221 is not limited to the above-described configuration, and may not have a shape bulging inward. That is, the inner wall 221 may be configured to have the same inner diameter without changing the inner diameter.

  As shown in FIG. 3, the laminated body 26 is configured by a two-layer structure of a net-like body 224 and a film 225, and these are bonded to each other.

  The net-like body 224 is constituted by, for example, a mesh structure or a net in which warp yarns and weft yarns are knitted, and the mesh is constituted by a diameter capable of passing the liquid L. However, it is necessary that the mesh 224 has a mesh size with a diameter that the above-described granular material 223 cannot pass through. Thereby, even if the liquid L passes through the mesh body 224, it is possible to prevent the passage of the granular body 223 and to hold the granular body 223.

  The film 225 is made of, for example, a resin, and is made of a material such as polyimide, polyethylene, polypropylene, polyester, polyvinyl chloride, or polycarbonate. The film 225 has a function of preventing the passage of the liquid L as well as the granular material 223.

  The net-like body 224 and the film 225 may be laminated by being bonded to each other, or may be laminated integrally. In addition, only the peripheral ends of the mesh 224 and the film 225 may be attached to each other. Moreover, the net-like body 224 and the film 225 are not limited to the case where they are laminated by being bonded to each other, and they may be separated from each other.

  Such a laminated body 26 is attached to the lower end of the lower chamber 220b in the container 220. As a result, the stacked body 26 serves as a bottom portion of the storage container 220. The planar form of the net-like body 224 and the film 225 constituting the laminated body 26 is in accordance with the planar view form of the lower chamber 220b to which it is attached.

  As shown in FIG. 4, the preprocessing main body 23 includes a cartridge housing portion 231 and a separation portion 233 extending downward from the cartridge housing portion 231. The separation unit 233 includes a first filter 234 and a second filter 235 provided downstream of the first filter 234, in other words, on the lower side. The lower end of the separation part 233 is continuous with the discharge port 236, and the discharge port 236 is normally sealed with a cap 237. The pretreatment body 23 may be made of any material such as glass, resin, metal, ceramics, and the like.

  The cartridge housing portion 231 is configured so that its periphery is surrounded by a peripheral wall 232, and the cartridge housing portion 231 has a shape in which the collection cartridge 22 can be inserted and attached.

  The separation part 233 is configured so that the inner diameter gradually decreases downward, but is not limited to this, and the inner diameters may all be the same. The separation part 233 has a cylindrical shape that is continuous from the cartridge housing part 231 and can guide the liquid to the discharge port 236 provided at the lower end thereof.

  In the separation unit 233, a first filter 234 is provided in order from the top, and a second filter 235 is provided at an interval from the first filter 234 toward the lower side.

  The first filter 234 is composed of a membrane capable of separating at least serum from the liquid L. That is, the serum separated from the liquid L is retained in the chamber R1 in the chamber R1 separated upward through the first filter 234 and the chamber R2 separated downward, and the serum is separated. It plays a role of transmitting L to the room R2.

  As the first filter 234, “Microza MF” manufactured by Asahi Kasei Chemicals Corporation, “Silicone Hollow Fiber” manufactured by Fuji Systems Co., Ltd., “Salto Clear Depth Filter 290-C4” manufactured by Sarutrius Stedum Biotech, etc. are used. May be. The first filter 234 is composed of a hollow fiber membrane, and serum is separated from the liquid L through pores formed between continuous tissues of the membrane.

  The second filter 235 is composed of a membrane capable of separating at least viruses and fungi from the liquid L. Specifically, the 2nd filter 235 is comprised with the film | membrane which can isolate | separate various viruses, such as Gram positive cocci, MRSA, SARS, and influenza virus. That is, in the chamber R2 that is separated upward through the second filter 235 and the chamber R3 that is separated downward, the virus and bacteria separated from the liquid L are retained in the chamber R2, and The separated liquid L is allowed to permeate into the chamber R3.

  As the second filter 235, “Microza UF” manufactured by Asahi Kasei Chemicals Corporation, “Salto Clear Depth Filter 290-F7H” manufactured by Sarutrius Stedum Biotech, or the like may be used. The configuration of the second filter 235 is not essential and may be omitted.

  The discharge port 236 is continuous from the chamber R3 in the separation unit 233, and the liquid L from which viruses and bacteria are separated is sent out. The discharge port 236 is formed at the tip protruding in a tubular shape toward the lower side, so that the convenience when the liquid L is injected toward the nucleic acid analysis chip 3 can be improved.

  The cap 237 is normally attached to the outlet 236 and seals it. When the liquid L is actually injected toward the nucleic acid analysis chip 3, the cap 237 is opened and the discharge port 236 is opened.

  Incidentally, the nucleic acid analysis chip 3 is not limited to the case where the collection cartridge 22 and the pretreatment main body 23 are configured to be separated from each other, but are configured to be integrated with each other. Also good. In addition, the configuration of the first filter 234 and the second filter 235 in the pretreatment main body may be omitted, and in such a case, at least only means for discharging the liquid from the collection cartridge 22 is mounted. That's fine. Specifically, the collection cartridge 22 and the pretreatment main body 23 are integrally configured, and this is realized by omitting the configuration of the first filter 234 and the second filter 235.

  Next, the configuration of the nucleic acid analysis chip 3 will be described. As shown in FIG. 5, the nucleic acid analysis chip 3 has an air pump connection port 31, a wash flow channel 32 continuous from the air pump connection port 31, a sample injection port 33, and a continuous sample from the sample injection port 33. A PCR channel 34, a heating wire 35 provided close to the PCR channel 34, an open / close switch 36 provided at the downstream end of the wash channel 32, and the wash channel 32 and the PCR channel 34 A hybridization channel 37 provided on the further downstream side where the downstreams are merged, a waste liquid storage reservoir 38 provided on the downstream side of the hybridization channel 37, and a discharge port 39 continuous to the waste liquid storage reservoir 38 It has.

  The air pump connection port 31 is connected to an air pump (not shown), and air is sent into the air pump connection port 31. The air and the cleaning liquid sent to the air pump connection port 31 are guided to the wash channel 32. The air pump connection port 31 may be closed by a lid (not shown) when not in use.

  The wash channel 32 is a channel for alternately flowing air and cleaning liquid. In order to flow such air and cleaning liquid alternately, a bifurcated branch air tube is attached to the entrance of the wash channel 32. And a washing | cleaning liquid is poured from one branched tube, and air is sent to the other branched through a micro pump from the tube. By increasing the pressure of the air sent from the micro pump, air enters between the cleaning liquids, and the air and the cleaning liquid flow alternately. This can also be realized by periodically changing the amount of air sent from the micropump and the atmospheric pressure.

  The interval between the alternately flowing air and the cleaning liquid may be any. The wash channel 32 is not limited to the case where air and cleaning liquid are alternately flowed. That is, the wash channel 32 may be a channel that allows only the cleaning liquid to flow. Note that the wash channel 32 may be omitted.

  The sample injection port 33 can be connected to the tip of the discharge port 236 in the pretreatment main body 23 described above. The liquid L is injected into the sample injection port 33 through the connected discharge port 236. The sample injection port 33 may be closed by a lid (not shown) when not in use. The sample injection port 33 is not necessarily limited to the case where the tip of the discharge port 236 in the preprocessing main body 23 can be connected. In such a case, the discharge port 236 in the pretreatment main body 23 may be positioned immediately above the sample injection port 33 while being separated from the sample injection port 33, and the liquid L may be dropped onto the sample injection port 33. That is, the sample injection port 33 may have any configuration as long as the liquid L can be fed from the discharge port 236 in the pretreatment main body 23. The liquid L sent to the sample injection port 33 is guided to the PCR channel 34.

  The PCR channel 34 amplifies the nucleic acid fragment in the target nucleic acid in the process of flowing the liquid L downstream. The PCR channel 34 may have any shape such as a circular shape or a polygonal shape in cross section, but it is desirable that the diameter of the PCR channel 34 be in the micron order of less than 1 mm. As a result, the liquid L passing through the PCR flow path 34 is in a so-called microfluidic state, and conversely, the PCR flow path 45 for flowing down such a microfluidic is in a so-called microflow path state. In addition, in order to allow the liquid L to flow efficiently downstream in a microfluidic state, the liquid channel L may be set so that its height gradually decreases as the PCR flow path 34 advances downstream.

  The PCR flow path 34 may have any path. For example, as shown in FIG. 5, the PCR flow path 34 is folded back so as to make a U-turn after going straight over a predetermined distance, and then again in a reverse direction. It may be folded back so as to make a U-turn after going straight over a distance, and this may be repeated. At this time, the number of U-turns may be set to any number. Such a terminal end of the PCR channel 34 joins the wash channel 32.

  In the present invention, instead of including a sterilizing component, a gene detection primer, and a polymerase enzyme in the liquid to be injected from the sample inlet 33, these substances are previously sprayed on the inner wall of the PCR channel 34 or the like. You may make it apply | coat. Thereby, it can be omitted that the liquid to be injected from the sample injection port 33 contains the sterilizing component, the gene detection primer, and the polymerase enzyme, and the liquid may be composed of water or the like. In addition, when a solvent composed of a liquid composed of water is injected from the sample injection port 33, the sterilizing component, gene detection primer, and polymerase enzyme applied on the inner wall of the PCR channel 34 are dissolved. Therefore, amplification of a desired nucleic acid fragment can be realized. In such a case, it is desirable that a turbulent flow is generated in the PCR flow path 34 and the structure can be stirred. Further, it is omitted that the sterilizing component, gene detection primer, and polymerase enzyme are contained in the liquid to be injected from the sample injection port 33, and these sterilization component, gene detection primer, and polymerase enzyme are individually supplied from the sample injection port 33. You may make it inject | pour.

  In the case where the substance in Table 1 or Table 2 is previously applied to the surface of the PCR flow path 34 and the PCR flow path 34 has a structure capable of causing turbulent flow and stirring, the solution L And the substance applied to the surface of the flow path may be dissolved and mixed. As a result, a process for detecting RNA by DNA amplification or cDNA synthesis can be performed in the PCR flow path 34.

  The heating wire 35 is provided below the PCR flow path 34. The heating wire 35 is extended so that its longitudinal direction is perpendicular to the straight direction of the PCR flow path 34. The heating wire 35 is set so that the heating temperature is 90 to 95 ° C. However, the heating temperature is not limited to this, and may be between 30 and 150 ° C. This heating wire 35 is not limited to the case where it is constituted by a single row, and may be constituted by a plurality of rows. In the example of FIG. 5, the width of the heating wire 35 is 5 mm, and the interval between the heating wires adjacent to each other is 5 mm. The PCR flow path 45 passing over the heating wire 35 is folded after passing in a direction orthogonal to the heating wire 35 provided at a predetermined interval, and further passed over each heating wire 35 and further folded. Thus, the passage over each heating wire 35 is repeated. As a result, after the PCR channel 34 is heated at 90-95 ° C. on the heating wire 35, it is cooled until passing through the next heating wire 35, and the temperature drops to about 55 ° C. As the next heating wire 35 approaches, the temperature rises and is heated to 90 to 95 ° C. In the process of flowing through the PCR channel 34, heating and cooling via the heating wire 35 occur repeatedly.

  The open / close switch 36 controls the cleaning liquid flowing from the wash flow channel 32 to the hybridization flow channel 37 by configuring the wash flow channel 32 to be openable and closable. As a specific configuration of the opening / closing switch 36, the wash channel 32 is formed of a resin tube, while only the position of the opening / closing switch 36 is formed of an elastically deformable film tube. An external pressure may be applied to the tube. By receiving such an external pressure, the tubular body made of film is elastically deformed so as to be reduced in diameter, whereby the passage of the cleaning liquid from the wash channel 32 can be suppressed. On the other hand, by preventing external pressure from being applied to the film tube, the film tube does not shrink, and in such a case, the washing solution passes through the open / close switch 36 and the hybridization channel 37. It will flow to. In order to apply the external pressure, for example, an air pump may be installed at the position where the opening / closing switch 36 is disposed, and the external pressure may be applied by supplying air to the tubular body made of film.

  The hybridization channel 37 has a nucleic acid probe formed in advance on the bottom surface. In the hybridization channel 37, a nucleic acid probe having a base sequence complementary to the target nucleic acid to be detected is formed in advance.

  In the hybridization channel 37, for example, when an analysis is performed based on a sample collected from excrement, the following bacteria in the intestinal flora (Bifidobacterium, lactic acid bacteria, Felicari bacteria, lean bacteria, Fraziris bacteria, butyricum bacteria, equol production Nucleic acid probes suitable for identifying bacteria, Welsh bacteria, Soldery bacteria, Nucleatum bacteria, Bacillus cereus, Copri bacteria, etc.) may be arranged.

  The hybridization channel 37 needs to be made of, for example, a light transmissive material so that light irradiated from the outside can reach and light emission can reach the outside. For example, it is made of a transparent plastic material.

  In the waste liquid storage reservoir 38, waste liquid containing nucleic acids other than the target nucleic acid flows from the hybridization channel 37. The waste liquid storage reservoir 38 is constituted by a water storage tank having a storage space in which the waste liquid can be temporarily stored.

  The discharge port 39 is provided for discharging the waste liquid accumulated in the waste liquid storage reservoir 38. The discharge port 39 is normally closed by a cap (not shown). By opening the cover, the waste liquid accumulated in the waste liquid storage reservoir 38 can be discharged.

  Next, the configuration of the chip measuring device 4 will be described. FIG. 6 is a system configuration diagram of the chip measuring device 4.

  The chip measuring device 4 includes an illumination unit 41 and a detection unit 42 connected to the control block 46, respectively, and a chip mounter 43 provided to face the illumination unit 41 and the detection unit 42. And a heat radiating plate 45 provided on the back side of the chip mounter 43. The heating wire 35 described above is attached to the chip mounter 43. A chip 3 for nucleic acid analysis is attached to the chip mounter 43 so as to be detachable.

  FIG. 7 shows a block configuration of the control block 46. The control block 46 includes a central control unit 61, a communication unit 62 connected to the central control unit 61, a display panel 63, an air pump control unit 64, a power stabilization unit 65, an operation unit 66, and a heating wire control. Part 67. The central control unit 61 is connected to the detection unit 42 and the illumination unit 41. An air pump 91 and a sensor 92 are connected to the air pump control unit 64. The heating wire 35 is connected to the heating wire control unit 67.

  The illumination part 41 is illumination which irradiates light with respect to the chip | tip 3 for nucleic acid analysis attached to the chip mounter 43, for example, is comprised by LED (blue LED etc.). The illumination unit 41 is arranged so that the irradiated light directly reaches the hybridization channel 37 in the nucleic acid analysis chip 3. In the illumination unit 41, the central control unit 61 controls ON / OFF of light emission, light emission intensity, light emission wavelength, light emission time, and the like.

  The detection unit 42 is a device for receiving a hybrid signal composed of light emitted from the nucleic acid analysis chip 3 attached to the chip mounter 43, converting it into an electrical signal, and transmitting it to the central control unit 61. is there. The detection unit 42 is composed of a solid-state imaging device such as a photomultiplier tube or a CCD image sensor. The detection unit 42 is disposed at a position where the light emitted from the hybridization channel 37 in the nucleic acid analysis chip 3 can be directly received. The detection unit 42 is controlled by the central control unit 61 for ON / OFF of light reception, light reception time, and the like.

  The detection unit 42 is not limited to receiving light in the visible light region, and may also measure light by receiving light in the infrared region, near infrared region, and ultraviolet region. In such a case, the detection unit 42 absorbs light in the infrared region and near infrared region from the hybridization channel 37 using, for example, a small spectroscope (C12666MA / C12800MA) manufactured by Hamamatsu Photonics. The molecular vibration may be measured by the method to identify the substance. Further, in order to detect minute light in the ultraviolet region or the like from the hybridization channel 37, the detection unit 42 is measured using a photomultiplier tube (eg, R11265U-200) of Hamamatsu Photonics. Also good.

  The chip mounter 43 is provided with a mounting table on which the nucleic acid analysis chip 3 can be mounted and a fixing device such as a magnet or an attachment so that the chip mounter 43 can be detachably fixed. The chip mounter 43 is configured such that the nucleic acid analysis chip 3 can be fixed so that the hybridization channel 37 is oriented in the direction of the illumination unit 41 and the detection unit 42.

  The heat radiating plate 45 is provided in the vicinity of the heating wire 35, and has a shape provided with irregularities that increase the outer surface area in order to easily radiate the heat generated from the heating wire 35. The heat radiating plate 45 is preferably made of a material that is lightweight and easy to process, such as aluminum.

  The heating wire control unit 67 performs feedback control so that the temperature of the heating wire 35 falls within a predetermined range under the control of the central control unit 61.

  The communication unit 62 controls the communication signal sent from the central control unit 61 to be converted into a radio signal and transmitted to the portable terminal 5 via the antenna. The communication unit 62 also receives a radio signal transmitted from the mobile terminal 5, converts it into an electric signal, and supplies it to the central control unit 61. The communication unit 62 is assumed to transmit and receive radio signals to and from the mobile terminal 5, but is not limited to this, and communicates with the public communication network 6 without using the mobile terminal 5. You may do it. The communication unit 62 is not limited to transmitting and receiving signals by wireless communication, and may be replaced by wired communication.

  The display panel 63 is composed of a liquid crystal display panel, for example, and is a device for displaying various information to the user.

  The operation unit 66 includes buttons, a mouse, a touch panel, and the like for the user himself to operate and control the chip measuring device 4. The input information input by the operation unit 66 is transmitted to the central control unit 61, and the central control unit 61 performs various controls based on the input information. Incidentally, when the operation unit 66 is configured by a touch panel, it may be configured integrally with the display panel 63.

  The power supply stabilization unit 65 performs control for stabilizing the power supply of the control block 46 and the heating wire 35 in the chip measuring device 4.

  The central control unit 61 is a central control unit for controlling the entire chip measuring apparatus 4 and includes, for example, a CPU (Central Processing Unit). The central control unit 61 transmits various control commands to each unit according to a user operation via the operation unit 66. The central control unit 61 controls the lighting unit 41 on / off of light emission, light emission intensity, light emission wavelength, light emission time, and the like. The central control unit 61 controls the detection unit 42 such as ON / OFF of light reception, light reception time, and the like. The central control unit 61 receives light reception information received from the hybridization channel 37 by the detection unit 42. The central control unit 61 displays such light reception information on the display panel 63 or transmits it to the outside via the communication unit 62. The central control unit 61 also performs open / close control by the open / close switch 36. The central control unit 61 executes each of these controls based on a program stored in a ROM (not shown) or the like.

  The air pump control unit 64 controls the air supply via the air pump 91 connected to the air pump connection port 31. The air supply amount and the air supply timing by the air pump control unit 64 may be controlled based on detection information by the sensor 92, for example.

  The sensor 92 includes, for example, a motion sensor, an infrared sensor, and the like, detects the movement of the user's hand, generates detection information based on the movement, and transmits the detection information to the air pump control unit 64. As a result, the air supply control of the air pump 91 by the air pump control unit 64 can be realized based on the movement of the user.

  The air pump 91 stores an air tank that stores air to be supplied by sucking air from the outside, and a tube that supplies the air stored in the air tank to the wash flow path 32 via the air pump connection port 31. And can be classified. The air pump 91 is not limited to the case of operating by automatic control, and air may be manually supplied to the wash channel 32 through the air pump connection port 31. In such a case, the configuration of the air pump control unit 64 and the sensor 92 can be omitted.

  Next, a method for analyzing a gene using the gene analysis system 1 to which the present invention is applied will be described.

  First, a user collects a sample collected from any one of a human body, excrement, and soil. The collection target here is blood, sweat, saliva and the like in the case of a human body, and feces and urine in the case of excrement.

  When collecting such a sample, the sample is attached to the tip of a rod-like body such as a cotton swab. Next, as shown in FIG. 8, the rod-like body 9 with the collected material 95 attached to the tip end portion 94 is filled in the collecting cartridge 22 in which a plurality of granular bodies 223 are filled in the container 220 and the liquid L is immersed. insert. The distal end portion 94 of the rod-shaped body 9 may be slightly thickened like a cotton swab, but is not limited thereto, and may be a rod-shaped body having the same diameter.

  As the rod-shaped body 9 is inserted into the receiving container 220 from the distal end portion 94, the distal end of the rod-shaped body 9 is directed toward the insertion port 229 through the inner diameter of the inner wall 221 that is gradually reduced in diameter from the upper end downward. It will be guided naturally. By pushing down the distal end portion 94 of the rod-shaped body 9 further downward from the insertion port 229, the distal end portion 94 is inserted into the insertion port 229 and enters the lower chamber 220b. At this time, a protruding portion 96 protruding outward may be formed in advance at a root portion slightly spaced from the distal end portion 94 of the rod-like body 9. As a result, the protrusion 96 is locked to the protrusion 222 in the insertion port 229, so that the pushing depth of the rod 9 can be controlled to an appropriate position. It can prevent that it pushes in too much and reaches the laminated body 26. FIG. Further, the rod-like body 9 may be previously formed with a crease, a kerf or the like so that it can be easily folded in the vicinity of the protruding portion 96 and easily separated from the distal end portion 94.

  The insertion port 229 formed by the inner wall 221 is biased in advance in the direction in which the inner diameter is narrowed, so that the distal end portion 94 of the rod-shaped body 9 having a large diameter is pushed into the insertion port 229. The operation of inserting the insertion port 229 to the lower chamber 220b side while pushing the insertion port 229 outward is performed. Then, when the thickened distal end portion 94 is completely in the lower chamber 220b side, the insertion port 229 that has been once expanded is urged toward the direction in which the inner diameter is narrowed again. The base portion at the distal end portion 94 of the rod-shaped body 9 is closed at the protruding portion 222 at the insertion port 229. As a result, it is possible to create a state in which the tip portion 94 cannot be directly visually recognized by the user. For this reason, if the collected material 95 is excrement, the user cannot directly see it, so that a clean impression can be given.

  Further, when the rod-shaped body 9 is pulled out from the lower chamber 220b, in the process of pushing out the thickened distal end portion 94 from the insertion port 229, the insertion port 229 is expanded outward and pulled toward the upper chamber 220a. Will be performed. At this time, if the sample 95 is attached to the distal end portion 94, the sample 95 can be dropped into the lower chamber 220b by applying the inner wall 221 that has received a biasing force to the sample 95. It becomes possible.

  Further, since the insertion port 229 formed by the inner wall 221 is biased in advance in the direction in which the inner diameter is narrowed, the lower chamber 220b is in a state close to hermetic sealing, so that the internal liquid L It becomes possible to make the granular material 223 last longer.

  Thus, the periphery of the tip 94 of the rod-shaped body 9 that has entered the lower chamber 220b is surrounded by the granular material 223. As a result, since the granular material 223 comes into contact with the tip portion 94, the sample 95 attached to the tip portion 94 also comes into contact with the granular material 223 in the same manner. As a result, the collected material 95 is wiped and collected by the contacted granular material 223. Since the granular material 223 is impregnated in the liquid L, the collected collected material is dissolved in the liquid L.

  Note that the user may agitate the rod-shaped body 9 in the lower chamber 220b or may slightly shake it up and down. As a result, the collected material 95 attached to the tip end portion 94 comes into contact with the granular material 223 several times and rubs, and the collected material 95 can be more effectively wiped off and dissolved in the liquid L.

  Next, the user pulls out the rod-shaped body 9 including the distal end portion 94 from which the collected matter 95 has been removed from the storage container 220. Next, as shown in FIG. 9, the sampling cartridge 22 is mounted in the cartridge housing portion 231 of the preprocessing main body 23. As a result, when the collection cartridge 22 and the pretreatment main body 23 are mounted, the storage container 220 filled with the granular material 223 and the first filter 234 are separated from each other via the stacked body 26. It becomes a state. In this state, the liquid L is held by the film 225 through which the liquid itself cannot pass.

  Next, a hole is made in the film 225 to cause the liquid L to flow down to the separation portion 233 side in the pretreatment main body 23. Any method may be used for making a hole in the film 225. For example, a plurality of long needles are installed upward in the chamber R1 separated upward via the first filter 234. The long needle pierces the film 225 when the sampling cartridge 22 is mounted, and the liquid L flows down from the hole opened in the film 225 to the separation portion 233 side of the pretreatment body 23. It becomes. On the other hand, since the granular body 223 cannot pass through the mesh body 224, the granular body 223 is not dropped into the separation part 233 by being held by the granular body 223.

  FIG. 10 shows a state where the liquid L is allowed to pass through the film 225 and flow down to the separation portion 233 side in the pretreatment main body 23. The liquid L first flows down into the chamber R1 and passes through the first filter 234. In the first filter 234, serum is separated from the liquid L through holes formed between the continuous tissues of the membrane. That is, the serum is accumulated in the chamber R1, and the liquid L from which the serum has been removed passes through the first filter 234 and flows down to the chamber R2.

  The liquid L that has flowed down into the chamber R2 is separated from at least viruses and fungi by the second filter 235. Only the liquid L from which the virus and bacteria have been separated passes through the second filter 235 and flows down to the chamber R3.

  After accumulating the liquid flowing down into the chamber R3, the user opens the cap 237 and connects the tip of the discharge port 236 in the pretreatment main body 23 to the sample injection port 33 in the nucleic acid analysis chip 3. As a result, the liquid L is injected into the sample injection port 33 through the discharge port 236 in the pretreatment main body 23. The liquid L sent to the sample injection port 33 is guided to the PCR channel 34.

  The liquid L guided to the PCR flow path 34 gradually flows down toward the downstream side of the flow path 34. At this time, the liquid L flows through the PCR channel 34 in a microfluidic state by flowing through the PCR channel 34 having a reduced diameter. The liquid L undergoes a polymerase chain reaction in the process of flowing through the PCR flow path 34. Specifically, when the PCR channel 34 flows on the heating wire 35, it is heated at 90 to 95 ° C., so at this stage, the nucleic acid is thermally denatured and the double-stranded nucleic acid Hydrogen bonds are broken and dissociated into single-stranded nucleic acids.

  The temperature decreases as the liquid L containing such single-stranded nucleic acid passes over the heating wire 35 and further away from the heating wire 35. Since this liquid L contains a primer, when the temperature is lowered to about 55 ° C., complementary binding to the primer occurs with respect to the single-stranded nucleic acid.

  Next, the temperature begins to rise again as the liquid L containing the single-stranded nucleic acid complementary to the primer is gradually brought closer to the heating wire 35. As the temperature rises, the primer begins to grow and becomes a double-stranded nucleic acid. Then, the liquid L reaches the heating wire 35 again and is heated at a temperature of 90 to 95 ° C. to cause denaturation, and the hydrogen bond of the double-stranded nucleic acid is cut and dissociated into the single-stranded nucleic acid. It will be.

  As the liquid L flows through the PCR channel 34, the above-described process occurs repeatedly. As a result, the nucleic acid fragment can be amplified and flow downstream.

  The liquid L exits the PCR channel 34 and is then introduced into the hybridization channel 37. In the stage where the liquid L flows through the hybridization channel 37, at least the hybridization channel 37 needs to reach the illumination light from the illumination unit 41, and light emission from the hybridization channel 37 occurs. In such a case, the central control unit 61 in the control block 46 needs to be controlled in advance so that the detection unit 42 can detect this.

  In the hybridization channel 37, if the target nucleic acid is contained in the liquid L, a stable duplex can be formed between this and the nucleic acid probe. As a result, only the target nucleic acid forms a duplex complementary to the nucleic acid probe, so that it remains in the hybridization channel 37, and any nucleic acid other than the target nucleic acid remains in the hybridization channel 37 as it is. It flows and is sent to the waste liquid storage reservoir 38.

  The presence or absence of the target nucleic acid bound to the nucleic acid probe remaining in the hybridization channel 37 is identified by the chip measuring device 4. The hybridization performed in the hybridization channel 37 may be performed by any known method. For example, when adopting fluorescence in situ hybridization, a nucleic acid probe labeled with a fluorescent substance or an enzyme is used, and a hybridization signal emitted by hybridization with a target nucleic acid is detected as a detection unit 42 as a fluorescence microscope. You may make it detect by.

  In this way, hybridization is performed in the hybridization channel 37. In order to perform hybridization more suitably in the hybridization channel 37, the temperature, pH, cation concentration and the like must be prepared in advance. The temperature may be adjusted using a Peltier element or a heating wire. For example, the temperature may be about 37 ° C. and the pH may be about 7.

  After completion of hybridization, the washing liquid may be flowed from the wash channel 32 to the hybridization channel 37 by opening the open / close switch 36. As a result, the nucleic acid analysis chip 3 can be used to identify the nucleic acid multiple times. When flowing the cleaning liquid from the wash flow path 32, the air pump 91 is operated via the air pump control unit 64 and the cleaning liquid is alternately supplied. The washing solution flows downstream through the wash channel 32 and then flows into the hybridization channel 37 as it is. The washing solution that has reached the hybridization channel 37 causes all remaining nucleic acid to flow and sends it to the waste solution storage reservoir 38.

  When the detection unit 42 can detect light emission indicating the presence of the target nucleic acid, the detection unit 42 outputs this to the central control unit 61. The central control unit 61 acquires the detection result and then displays the detected data via the display panel 63 or transmits the detected data to the mobile terminal 5 via the communication unit 62.

  When the mobile terminal 5 receives the data via the communication unit 62, the mobile terminal 5 uses the examination application stored in the mobile terminal 5 to analyze the data, identify the nucleic acid, and perform the gene analysis. . The form terminal 5 displays the analysis result to the user or displays various proposals for future health maintenance to the user. The mobile terminal 5 may further transmit the data received via the communication unit 62 to the public communication network 6. The data transmitted to the public communication network 6 is stored in the server 7. The data stored in the server 7 is analyzed by the central control device 8 and confirmed by a doctor or the like. Further, knowledge obtained by analyzing information by the central control device 8 and analysis results may be transmitted to the portable terminal 5 via the public communication network 6.

  According to the present invention having the above-described configuration, the user collects a sample from the human body, excrement, or soil with the rod-shaped body 9, and collects the sample at the distal end portion 94 in the collection cartridge 22 in which the liquid L is immersed. The rod-like body 9 to which the object 95 is attached is inserted. In addition, the operation of attaching the collection cartridge 22 to the pretreatment main body 23 and the connection of the tip of the discharge port 236 in the pretreatment main body 23 to the sample injection port 33 of the nucleic acid analysis chip 3 All will be automatically analyzed.

  For this reason, even a user who has no specialized knowledge about medical care can perform genetic analysis by himself with very simple work.

  In addition, since the pretreatment kit 2 for gene analysis, the chip 3 for nucleic acid analysis, and the chip measuring device 4 are not complicated, the size of the device can be reduced and it is suitable for self-examination at home. It can be purchased even by an individual because it is structured and can be provided at low cost.

  Furthermore, according to the present invention, it is possible to extract nucleic acid from a collected sample with high efficiency, and in order to identify nucleic acid with high accuracy, serum, bacteria, and viruses are removed in advance through the pretreatment kit 2 for gene analysis. In addition, these operations can be performed very easily without requiring special skills and knowledge.

  In addition, this can be washed by flowing a washing solution from the wash channel 32 to the hybridization channel 37, and the nucleic acid analysis chip 3 can be used over and over again. The cleaning operation can be performed very simply.

  In addition, while simplifying the device configuration as described above, improving ease of handling, and improving cost performance, the accuracy of biochemical tests can be maintained at the same level as before, so the same level at medical institutions. Can be realized at home.

  The present invention is not limited to the embodiment described above. You may comprise the granular material 223 by enclosing the liquid L beforehand in the skin which consists of elastic substances. In such a case, it is not essential to impregnate the liquid L in the storage container 220. When the rod-like body 9 with the collected material attached to the tip is inserted, the collected material is collected by bringing the tip into contact with the granular material 223 as described above. In addition to this, the liquid L is caused to flow out by breaking the skin of the granular body 223 by the tip of the rod-shaped body 9. As a result, the tip of the rod-shaped body 9 can be impregnated with the liquid L that has flowed out.

  Moreover, the gene analysis system 1 to which the present invention is applied may be configured as a case accommodation type as shown in FIGS. Since the components and members of the case-accommodating gene analysis system 1 ′ are the same as those in the above-described embodiment, the same reference numerals are used for the description, and the following description is omitted.

  FIG. 11 shows the appearance of the case-accommodating gene analysis system 1 ′. In the case-accommodating gene analysis system 1 ', one end is joined by a hinge (not shown), and the case 100a and the case 100b are configured to be openable and closable around the hinge (not shown). A handle 101 is provided on the upper surface of the cases 100a and 100b, and a lock mechanism 102 is provided. A display 103 and a touch panel 104 may be provided outside the case-accommodating gene analysis system 1 ′. Incidentally, the display 103 corresponds to the display panel 63, and the touch panel 104 corresponds to the operation unit 66.

  FIG. 12 shows a state in which the cases 100a and 100b are developed. The above-described chip measuring device 4 is disposed on the case 100 a side, and a personal computer (PC) 50 is connected to the chip measuring device 4, and a battery 105 is connected to the PC 50. The PC 50 operates with power supplied from the battery 105. A measurement result by the chip measuring device 4 is sent to the PC 50. This PC 50 bears the same function as the portable terminal 5 described above.

  A nucleic acid analysis chip 3 is disposed on the case 100b side, and a pretreatment body 23 is connected to the nucleic acid analysis chip 3. The nucleic acid analysis chip 3 is provided at a position just facing the chip measurement device 4 when the cases 100a and 100b are closed. Thereby, the measurement mentioned above is attained. A discharge port 236 in the pretreatment main body 23 is directly connected to the sample injection port 33 in the nucleic acid analysis chip 3 so that the liquid L is sent. The pretreatment main body 23 and the nucleic acid analysis chip 3 may be arranged in a plurality of sets. The pretreatment main body 23 can introduce a sample from the sample introduction port 108 via the branch tube 111. The configuration of the sample introduction port 108 is the same as that of the collection cartridge 22. The liquid L from the collection cartridge 22 is sent to the pretreatment body 23 through the branch tube 111. Incidentally, the power for feeding the liquid L from the collection cartridge 22 to the branch tube 111 is assumed to be performed via the pump 109. The inside of the cases 100a and 100b is covered with a reinforced plastic or a reinforced protective film except for the sample introduction port 108, so that the specimen can be prevented from scattering.

  According to such a case-accommodating gene analysis system 1 ′, the system can be compactly assembled into a case having a size such as a suitcase. In addition, high-risk viruses and illnesses (HIV, Marburg virus, Ebola virus, smallpox, Lassa fever, etc.) can be measured quickly without entrusting them to special management facilities. It is possible to quickly detect trends and suppress damage expansion. In particular, according to this case-accommodating type gene analysis system 1 ′, the rod-like body 9 to which the collected material 95 is attached can be measured automatically only by inserting it into the sample introduction port 108. In addition, since it can be disposed of without touching human hands from the start to the end of measurement, it is excellent in terms of hygiene.

  Moreover, according to the present invention, in addition to nucleic acid measurement, it is possible to measure biological substances such as peptides, exosomes, biomarkers, and antibodies in the same manner. In such a case, the liquid L even injected through the discharge port 236 in the pretreatment main body 23 will flow directly into the hybridization channel 37 without performing nucleic acid amplification in the PCR channel 34. Other processes are the same as those in the above-described embodiment. That is, when measuring biological substances such as peptides, exosomes, biomarkers, and antibodies in the same manner, as an alternative to the PCR flow path 34, an injection flow path for flowing the injected liquid L to the hybridization flow path 37; It only has to be done.

DESCRIPTION OF SYMBOLS 1 Gene analysis system 2 Gene analysis pretreatment kit 3 Nucleic acid analysis chip 4 Chip measuring device 5 Portable terminal 6 Public communication network 7 Server 8 Central control device 9 Rod 12 Public communication network 21 Plunger 22 Collection cartridge 23 For preprocessing Main body 26 Laminate 31 Air pump connection port 32 Wash flow channel 33 Sample injection port 34 PCR flow channel 35 Heating wire 36 Open / close switch 37 Hybridization flow channel 38 Waste liquid storage reservoir 39 Discharge port 41 Illumination unit 42 Detection unit 43 Chip mounter 45 Heat dissipation Plate 45 Flow path 46 Control block 61 Central control unit 62 Communication unit 63 Display panel 64 Air pump control unit 65 Power supply stabilization unit 66 Operation unit 67 Heating wire control unit 91 Air pump 92 Sensor 95 Sample 96 Projection part 211 Inner cylinder part 212 Pusher 213 Rubber part 220 Container 2 0a upper chamber 220b lower chamber 221 inner wall 222 the protrusion 223 granulate 224 mesh body 225 film 229 insertion opening 231 cartridge accommodating portion 232 a peripheral wall 233 separating unit 234 first filter 235 second filter 236 outlet 237 cap

Claims (17)

In a pretreatment kit for gene analysis for extracting biological material from a sample collected from human body, excrement, or soil,
When a rod-shaped body in which a plurality of granular bodies are filled and a liquid is immersed in the granular body and the collected material is attached to the tip is inserted, the tip is brought into contact with the granular body. A collecting cartridge for collecting the collected material and impregnating the liquid;
A pretreatment kit for gene analysis, comprising: discharge means for discharging the liquid delivered from the collection cartridge to the outside. A first filter for separating at least serum from the liquid delivered from the attached collection cartridge, and a pretreatment main body having the discharge means;
The pretreatment kit for gene analysis according to claim 1, wherein the discharge means discharges the liquid that has passed through the first filter to the outside.   The pretreatment kit for gene analysis according to claim 1 or 2, wherein the liquid contains a bactericidal component, a gene detection primer, and a polymerase enzyme. The main body for pretreatment further includes a second filter for separating at least viruses and bacteria from the liquid that has passed through the first filter, and the outlet has the liquid that has passed through the second filter as an outside. The pretreatment kit for gene analysis according to claim 3, wherein the pretreatment kit is for gene analysis. The sampling cartridge is pre-biased in a narrowing direction and has an insertion port into which the tip of the rod-shaped body is inserted, and the granular material is filled in the insertion port. The pretreatment kit for gene analysis according to any one of claims 1 to 4, wherein the liquid is immersed therein. 6. The rod-shaped body according to claim 5, wherein the rod-shaped body has a protrusion that protrudes outward from a root portion of a tip to which the collected material is attached, and the protrusion is locked to the insertion port. Pretreatment kit for gene analysis. When the collection cartridge and the pretreatment main body are mounted, the storage container filled with the granular material and the first filter are a net-like body through which the liquid can pass and a film through which the liquid cannot pass. Are separated by a stack of layers,
The liquid is supplied from the sampling cartridge, and a hole for allowing the liquid to pass therethrough is formed in the film so that the liquid is allowed to pass therethrough and the granular material is held by the mesh. The pretreatment kit for gene analysis according to any one of 1 to 6. The granule is made of any one of metal, ceramics, and resin, and any one of zeolite, emulsion gel, hydrophilic polymer, and amino acid substance is formed on the surface thereof. The pretreatment kit for gene analysis according to any one of Items 1 to 7. In a pretreatment kit for gene analysis for extracting biological material from a sample collected from human body, excrement, or soil,
The epidermis is filled with a plurality of granular materials composed of an elastic material, and a liquid containing a bactericidal component, a gene detection primer, and a polymerase enzyme is enclosed in the epidermis, and the collected material is attached to the tip. A collecting cartridge that impregnates the liquid that has flowed out by breaking the epidermis with the tip when the tip is brought into contact with the granular body when the rod is inserted ,
A pretreatment kit for gene analysis, comprising: discharge means for discharging the liquid delivered from the collection cartridge to the outside. In a nucleic acid analysis chip for analyzing a target nucleic acid to be detected from a sample collected from a human body, excrement, or soil,
A PCR flow channel in which a liquid containing a biological substance extracted from a primer for gene detection, a polymerase enzyme, and the collected material is injected, and a nucleic acid fragment in the target nucleic acid is amplified and flowed downstream;
A hybridization channel that is continuous from the PCR channel and that allows the amplification product flowing from the PCR channel to hybridize while flowing downstream;
A nucleic acid analysis chip comprising: target nucleic acid analysis means for identifying the target nucleic acid by measuring a hybridization signal generated in the hybridization channel. It further includes a wash channel for flowing at least the cleaning liquid,
11. The hybridization channel is continuous from the wash channel, and can be cleaned by flowing a cleaning solution from the wash channel after the hybridization. Chip for nucleic acid analysis. The PCR channel is configured to have a diameter on the order of microns, and is disposed so as to pass over a heating wire having a heating temperature of 30 to 150 ° C. provided at a predetermined interval. The nucleic acid analysis chip according to claim 10 or 11, wherein the nucleic acid is heat denatured. The PCR flow path is arranged so as to be repeated after passing over the heating wire provided at a predetermined interval, and further passing over the heating wire, and further passing back over the heating wire. The chip for nucleic acid analysis according to claim 12. 12. The nucleic acid analysis chip according to claim 11, wherein the wash channel and the washing liquid and air flow alternately. The nucleic acid analysis chip according to any one of claims 10 to 14, further comprising communication means for transmitting information detected by the target nucleic acid analysis means to the outside via a communication network. A pretreatment kit for gene analysis according to any one of claims 1 to 9,
A nucleic acid analysis chip according to any one of claims 10 to 15,
A gene analysis system, wherein the liquid discharged from the discharge port in the gene analysis pretreatment kit flows into a PCR flow path in the nucleic acid analysis chip. In a biological material analysis chip for analyzing a biological material to be detected from a sample collected from a human body, excrement, or soil,
An injection flow path in which a liquid containing a biological material extracted from the collected material is injected, and this flows downstream;
A hybridization flow channel that is continuous from the injection flow channel and allows the biological material flowing from the injection flow channel to hybridize while flowing downstream;
A target nucleic acid analysis means for identifying the target nucleic acid by measuring a hybridization signal generated in the hybridization channel;
At least a wash channel for flowing the cleaning liquid is provided,
The hybridization channel is continuous from the wash channel, and can be washed by flowing a washing solution from the wash channel after the hybridization. Chip.

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