JP6982338B2 - Pretreatment kit for gene analysis, chip for nucleic acid analysis, analysis system, chip for biomaterial analysis - Google Patents

Description

The present invention is a pretreatment kit for gene analysis for extracting a biological substance from a collection collected from the human body, excrement, or soil, and a nucleic acid analysis for analyzing a target nucleic acid to be detected from the collection. It relates to a chip, an analysis system and a chip for biomaterial analysis.

Traditionally, medical examinations have been conducted exclusively at specialized medical institutions such as hospitals, but with the advent of a super-aging society in Japan, the burden of examinations at medical institutions has become seriously increasing. For this reason, tests that require a dedicated device such as X-ray photography and electrocardiogram measurement, and tests that require a syringe such as blood tests will continue to be performed at medical institutions, while users will be able to perform simple tests. If possible, it is also necessary to consider reducing the burden of examinations at medical institutions by using self-examinations conducted at home together. 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 provided, for example.

Under these circumstances, in order to enable next-generation self-examination, in addition to downsizing, generalization, and price reduction of examination equipment, even users who do not have specialized knowledge about medical treatment can realize it. A degree of ease of handling is required.

In recent years, gene analysis using a nucleic acid analysis chip (DNA chip) has been realized in response to such social needs. Chips for nucleic acid analysis are used in various fields such as medical field, chemical field, drug discovery field, clinical testing field, food field, agricultural field, engineering field, forensic medicine field and criminal identification field, for example, medical field. In general, various diseases such as cancer are screened by a test using a biochemical sample such as blood.

A nucleic acid analysis chip is a device in which a nucleic acid probe is densely immobilized on a glass or silicon substrate. When performing gene analysis with high accuracy using a nucleic acid analysis chip, gene amplification is required in advance. The nucleic acid thus amplified will then bind to, in the hybridization step, if it has a complementary base sequence with the nucleic acid probe on the substrate. 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 the DNA samples that did not bind to the nucleic acid probe on the substrate are washed away.

Subsequently, the presence / absence 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 a fluorescent substance 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 the analysis process can be performed based on the measured amount.

In the nucleic acid analysis chip as described above, for example, the technique shown in Patent Document 1 is disclosed for the purpose of meeting the needs for self-examination performed at home. Patent Document 1 discloses a nucleic acid analysis chip which can detect a reaction instantly, has a compact 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.

Japanese Unexamined Patent Publication No. 2004-28589 Japanese Patent Publication No. 2014-52883

However, in the conventional nucleic acid analysis chip described above, there is room for further improvement from the viewpoint of improving the accuracy of the biochemical test. In addition, when the nucleic acid analysis chip is fixed to the analyzer to perform an enzymatic reaction or a biochemical reaction, it may be difficult to control the temperature of the nucleic acid analysis chip with high accuracy, and the desired reaction may not proceed sufficiently. there were.

Patent Document 1 proposes that a nucleic acid analysis chip is provided with a suction groove, and the chip is sucked and fixed at a desired position by vacuum suctioning through the suction groove with a vacuum exhaust pump. However, if such a configuration is adopted, it becomes a big obstacle in reducing the size of the analyzer.

Therefore, the present invention has been devised in view of the above-mentioned problems, and an object of the present invention is to reduce the size and generalization of a testing device in a nucleic acid analysis chip that enables next-generation self-diagnosis. In addition to lowering the price, the ease of handling that can be realized even by users without specialized knowledge about medical care is ensured, and the accuracy of biochemical tests can be further improved. Nucleic acid analysis chip, gene analysis It is an object of the present invention to provide a pretreatment kit for gene analysis for extracting a biological substance from a collection collected from a human body, excrement, or soil in the stage before performing the system and its analysis.

The pretreatment kit for gene analysis according to the first invention is a pretreatment kit for gene analysis for extracting a biological substance from a collection collected from any of the human body, excrement, and soil, and the epidermis is composed of an elastic substance. A liquid containing a bactericidal component, a primer for gene detection, and a polymerase enzyme was encapsulated in the epidermis, and a rod-shaped body having the collected material attached to the tip was inserted. In this case, the sample is collected by contacting the tip with the granules, and the collection cartridge is impregnated with the liquid that has flowed out by breaking the epidermis with the tip, and the collection cartridge is sent out. Pretreatment having the discharge means for discharging the liquid to the outside, the first filter for separating at least serum from the liquid sent out from the attached collection cartridge, and the discharge means. The discharge means discharges the liquid that has passed through the first filter to the outside, and is filled with the granules when the collection cartridge and the pretreatment main body are mounted. The storage container and the first filter are separated from each other by 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 to each other, and when the liquid is delivered from the collection cartridge. It is characterized in that the film is formed with holes for passing the liquid to allow the liquid to pass through, and the granules are held by the reticulated body .

The analysis system according to the second invention is a pretreatment kit for gene analysis according to the first invention, and a nucleic acid analysis for analyzing a target nucleic acid to be detected from a sample collected from any of the human body, excreta, and soil. A PCR flow path in which a liquid containing a gene detection primer, a polymerase enzyme, and a biological substance extracted from the above-mentioned sample is injected, and the nucleic acid fragment in the target nucleic acid is amplified and flows downstream, and the above-mentioned PCR flow. The target nucleic acid is measured by measuring the hybridization signal that flows downstream from the amplification product that is continuous from the pathway and flows in from the PCR flow path, and the hybridization signal generated in the hybridization flow path. A nucleic acid analysis chip having a target nucleic acid analysis means for identifying the nucleic acid is provided, and the liquid discharged from the discharge port in the gene analysis pretreatment kit is allowed to flow into the PCR flow path in the nucleic acid analysis chip. It is a feature.

In the second invention, the analysis system according to the third invention further includes at least a wash flow path through which the washing liquid flows, and the hybridization flow path is continuous from the wash flow path, and after the hybridization, the wash flow is performed. It is characterized in that it can be washed by flowing in the cleaning liquid from the road.

In the analysis system according to the fourth invention, in the second invention or the third invention, the above-mentioned PCR flow path is composed of a diameter on the order of micron, and electric heat provided at predetermined intervals and having a heating temperature of 30 to 150 ° C. It is arranged so as to pass on the wire, and is characterized by thermally denaturing the target nucleic acid when passing on the heating wire.

In the fourth aspect of the invention, the analysis system according to the fifth invention is the present invention, in which the PCR flow path is passed over a heating wire provided at a predetermined interval, then folded back, further passed over the heating wire, and further folded back to be further heated. It is characterized in that it is arranged so as to repeatedly pass on the line.

The analysis system according to the sixth aspect of the invention is characterized in that, in the third aspect of the invention, the washing liquid and air flow alternately in the wash flow path.

The analysis system according to the seventh invention is characterized in that, in any one of the second invention to the sixth invention, the communication means for transmitting the information detected by the target nucleic acid analysis means to the outside via the communication network is further provided. do.

According to the present invention having the above-described configuration, the user collects a sample from any of the human body, excrement, and soil with a rod-shaped body, and the sample is attached to the tip of the collection cartridge in which the liquid is immersed. Insert the rod-shaped body. Other than that, all you have to do is attach the collection cartridge to the pretreatment body, and connect the tip of the discharge port in the pretreatment body to the sample injection port of the nucleic acid analysis chip, and all the rest will be done automatically. Analysis will be carried out.

Therefore, even a user who does not have specialized knowledge about medical treatment can perform gene analysis by himself / herself with an extremely simple task.

In addition, since the pretreatment kit for gene analysis, the chip for nucleic acid analysis, and the chip measuring device are not complicated in device configuration, they can be miniaturized and have a structure suitable for self-examination at home. In addition, since it can be provided at a low price, it can be purchased by individuals.

Further, according to the present invention, nucleic acid can be extracted from the collected sample with high efficiency, and serum, bacteria, and viruses are removed in advance through a pretreatment kit for gene analysis in order to identify the nucleic acid with high accuracy. Moreover, these operations can be performed extremely easily without the need for special skill or knowledge.

In addition, it can be washed by flowing the washing liquid from the washing flow path to the hybridization flow path, and the nucleic acid analysis chip can be used repeatedly, so that the configuration is very excellent in cost performance. Therefore, this cleaning work can be performed extremely easily.

In addition, while improving the device configuration, ease of handling, and cost performance as described above, the accuracy of biochemical tests can be maintained at the same level as before, which is the same level at medical institutions. It will be possible to realize the gene analysis of the above at home.

It is a figure which shows the system structure of the gene analysis system to which this invention was applied. It is a figure for demonstrating the structure of the collection cartridge. It is a figure for demonstrating the details of a laminated body composed of a net-like body and a two-layer structure of a film. It is a figure for demonstrating the structure of the main body for preprocessing. It is a figure for demonstrating the structure of the chip for nucleic acid analysis. It is a system block diagram of a chip measuring device. It is a figure which shows the block composition of the control block in a chip measuring apparatus. It is a figure for demonstrating the method of analyzing a gene using the gene analysis system to which this invention is applied. It is a figure which shows the state which attached the collection cartridge to the cartridge accommodation part in the pretreatment main body. It is a figure which shows the state which the film passed through the liquid and flowed down to the separation part side in the main body for pretreatment. It is a figure which shows the appearance of the case-contained gene analysis system. It is a development view of the case-contained gene analysis system.

Hereinafter, a mode 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 can communicate with the pretreatment kit 2 for gene analysis, the nucleic acid analysis chip (DNA chip) 3, the chip measuring device 4 to which the nucleic acid analysis chip 3 is mounted, and the chip measuring device 4. The mobile terminal 5 is provided with a server 7 connected to the mobile terminal 5 via the public communication network 6, and a central control device 8 connected to the 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 a public communication network 6 at least based on a user's operation. be. The mobile terminal 5 is a device that can be carried at all times by making it portable by the user, but the present invention is not limited to this, and includes all electronic devices such as desktop PCs.

The mobile terminal 5 can download the examination application via the public communication network 6. As a result, the user can activate the examination application by operating the mobile terminal 5 to check his / her own examination record or to confirm various proposals for maintaining health in the future. In the following example, the 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 a mobile terminal 5, a server 7, and a central control device 8 are connected via a communication line. The public communication network 6 may be configured by a so-called optical fiber communication network. Further, the public communication network 6 is not limited to the wired communication network, and may be realized by a wireless communication network.

A predetermined database is constructed on 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 the request from the mobile terminal 5.

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

The pretreatment kit 2 for gene analysis 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 portion 211 is extended in the longitudinal direction, and a rubber portion 213 is provided at the extended end thereof. The pusher 212 is provided on the other end side of the plunger 21 different from the extension end where the rubber portion 213 is provided, and the inner cylinder portion 211 is collected by pressing the user's finger against the cartridge. It is configured to be freely insertable in 22.

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

The granular material 223 may be made of any material such as resin, ceramics, and metal. The granular body 223 may be made of die-cast when it is made of metal, or may be made of polystyrene, for example, when it is made of resin. Further, in the granular material 223, these materials may be laminated in a two-layer structure, and for example, a resin layer may be coated on the outer shell of a metallic core made of die-cast.

The granular body 223 will be described by taking the case of being composed of spherical beads as an example, but the present invention is not limited to this, and the granular body 223 may be composed of any polyhedron including a cube and a rectangular parallelepiped. , May be composed of random three-dimensional shapes. Further, the granular body 223 may be formed of a specific shape such as a hook shape.

The surface of the granular material 223 may be further coated with calcium phosphate. Through this layer of calcium phosphate, it is possible to make the pH of the container 220 approach neutral. Further, by coating the surface of the granular material 223 with any substance such as zeolite, emulsion gel, hydrophilic polymer, or amino acid substance, stool or solid matter as excrement is adsorbed on the granular material 223. It is possible to make it easier to get rid of extra things with a filter.

The liquid L is pre-injected into the storage container 220 filled with the granular material 223 having the above-described configuration. As a component of this liquid, for example, if the sample to be extracted from the genomic DNA as a biological substance is stool, it may be composed of the composition shown in Table 1 below. Further, in the case of extracting RNA, the liquid may be composed of the composition shown in Table 2. The volume of each component composition shall be expressed in% by weight with respect to the total weight of the liquid L.

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

If the target collection is blood or urine, it may be composed of the compositions shown in Table 3 below.

Further, in the case of extracting RNA, the liquid may be composed of the composition shown in Table 4.

Here, the adjusting buffer is a stabilizing substance for preventing blood coagulation, and is composed of EDTA, SDS and the like. The buffer solution is a solution for preventing decomposition and inactivation of genomic DNA, and is composed of a mixture of citric acid and disodium hydrogen phosphate. The DNase / RNase-treated water is water from which DNase and the like, which are DNA-degrading enzymes, have been removed, and is composed of DNA-degrading enzyme and water from which RNA-degrading enzyme has been removed.

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

Further, in the case of extracting RNA, the liquid may be composed of the composition shown in Table 6.

Here, the buffer solution is a solution for preventing decomposition and inactivation of genomic DNA, and is composed of a mixture of citric acid and disodium hydrogen phosphate. The DNase / RNase-treated water is water from which DNase and the like, which are DNA-degrading enzymes, have been 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, hydroxypropyl cellulose, pronase and the like.

The inner wall 221 is made of an elastic body such as rubber, and is elastically configured to expand and contract in diameter in the radial direction of the storage container 220. The inner diameter of the inner wall 221 is gradually reduced from the upper end to the lower side of the storage container 220, and the inner diameter thereof becomes the smallest in the middle stage. In the middle stage where the inner diameter is the smallest, an insertion port 229 surrounded by a protrusion 222 that further protrudes inward is provided. The inner diameter thereof is formed so as to gradually increase downward from the insertion port 229. As a result, the storage 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 inner cylinder portion 211 of the plunger 21 described above can be inserted into the upper chamber 220a. The lower chamber 220b is filled with the granular material 223, and the liquid L is further injected. The granules 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 preliminarily urged in a direction in which the inner diameter thereof is narrowed. That is, the insertion slot 229 may be preliminarily urged in the direction of the arrow in FIG.

Incidentally, the configuration of the inner wall 221 is not limited to the configuration described above, and may not have a shape that bulges inward. That is, the inner diameters of the inner walls 221 may be configured to be the same without changing.

As shown in FIG. 3, the laminated body 26 is composed of a two-layer structure of a network body 224 and a film 225, and these are attached to each other.

The mesh body 224 is composed of, for example, a mesh structure or a mesh in which warp threads and weft threads are woven, and the mesh of the mesh is configured with a diameter that allows the liquid L to pass through. However, the net-like body 224 needs to have a mesh having a diameter such that the above-mentioned granular body 223 cannot pass through. As a result, even if the liquid L passes through the reticulated body 224, it is possible to prevent the granular body 223 from passing through and to retain 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 network body 224 and the film 225 may be laminated by being adhered to each other, or may be integrated and laminated. Further, only the peripheral ends of the net-like body 224 and the film 225 may be attached to each other. Further, the net-like body 224 and the film 225 are not limited to the case where they are laminated by being adhered to each other, and they may be separated from each other.

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

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

The cartridge accommodating portion 231 is configured to be surrounded by a peripheral wall 232, and has a shape in which a collecting cartridge 22 can be inserted and mounted.

The separating portion 233 is configured so that the inner diameter gradually decreases downward, but the inner diameter is not limited to this, and all the inner diameters may be the same. The separation portion 233 has a tubular shape that is continuous from the cartridge accommodating portion 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 intervals 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, in the chamber R1 separated to the upper side via the first filter 234 and the chamber R2 separated to the lower side, the serum separated from the liquid L is retained in the chamber R1 and the serum is separated. It plays a role of transmitting L to the chamber R2.

For the first filter 234, "Microza MF" manufactured by Asahi Kasei Chemicals Co., Ltd., "Silicone hollow thread" manufactured by Fuji Systems Co., Ltd., "Zart Clear Depth Filter 290-C4" manufactured by Sarutris Stedum Biotech, etc. are used. You may. The first filter 234 is composed of a hollow fiber membrane and separates serum from the liquid L through pores formed between the continuous tissues of the membrane.

The second filter 235 is composed of a membrane capable of separating at least viruses and bacteria from the liquid L. Specifically, the second filter 235 is composed of a membrane capable of separating various viruses such as Gram-positive cocci, MRSA, SARS, and influenza virus. That is, in the chamber R2 separated to the upper side via the second filter 235 and the chamber R3 separated to the lower side, the virus and the bacterium separated from the liquid L are retained in the chamber R2, and the virus and the bacterium are accumulated. It plays a role of allowing the separated liquid L to permeate the chamber R3.

As the second filter 235, "Microza UF" manufactured by Asahi Kasei Chemicals Co., Ltd., "Zart Clear Depth Filter 290-F7H" manufactured by Sarutrias 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 the virus and the bacterium are separated is sent out. Since the discharge port 236 is formed at the tip protruding downward in a tubular shape, the convenience of injecting the liquid L toward the nucleic acid analysis chip 3 can be improved.

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

Incidentally, in the nucleic acid analysis chip 3, the collection cartridge 22 and the pretreatment main body 23 are not limited to the case where they are separated from each other, and these are integrated with each other. It is also good. Further, the configuration of the first filter 234 and the second filter 235 may be omitted in the pretreatment main body, and in such a case, at least a means for discharging the liquid from the collection cartridge 22 is mounted. Just do it. Specifically, this can be realized by integrally configuring the collection cartridge 22 and the pretreatment main body 23, and omitting the configurations 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 is continuous from the air pump connection port 31, the wash flow path 32 continuous from the air pump connection port 31, the sample injection port 33, and the sample injection port 33. The PCR flow path 34, the heating wire 35 provided close to the PCR flow path 34, the open / close switch 36 provided at the downstream end of the wash flow path 32, and the wash flow path 32 and the PCR flow path 34. A hybridization flow path 37 provided on the downstream side where each downstream merges, a waste liquid storage reservoir 38 provided on the downstream side of the hybridization flow path 37, and a discharge port 39 continuous with the waste liquid storage reservoir 38. It is equipped with.

An air pump (not shown) is connected to the air pump connection port 31, and air is sent to 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 flow path 32. The air pump connection port 31 may be closed by a lid (not shown) when not in use.

The wash flow path 32 is a flow path for alternately flowing air and a cleaning liquid. In order to allow such air and cleaning liquid to flow alternately, a bifurcated air tube is attached to the entrance of the wash flow path 32. Then, the cleaning liquid is flowed from one of the branched tubes, and air is sent from the tube to the other branched via a micropump. By increasing the air pressure of the air sent from the micropump, air enters between the cleaning liquids, and the air and the cleaning liquid flow alternately. This can also be achieved by periodically fluctuating the amount and pressure of air sent from the micropump.

The interval between the alternating air and the cleaning liquid may be any. The wash flow path 32 is not limited to the case where air and a cleaning liquid are alternately flowed. That is, the wash flow path 32 may be one in which only the cleaning liquid flows. The wash flow path 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 pretreatment main body 23 can be connected. In such a case, the discharge port 236 of the pretreatment main body 23 may be positioned directly above the sample injection port 33 in a state of 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 be any structure as long as the liquid L can be sent 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 flow path 34.

The PCR flow path 34 amplifies the nucleic acid fragment in the target nucleic acid in the process of flowing the liquid L downstream. The cross-sectional shape of the PCR flow path 34 may be any shape including a circular shape and a polygonal shape, but it is desirable that the diameter thereof is on the order of microns 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 through which such a microfluidic flows is in a so-called microfluidic state. In addition, in order to allow the liquid L to efficiently flow downstream in the state of a microfluidic, the height may be set to gradually decrease as the PCR flow path 34 advances downstream.

The PCR flow path 34 may be any path, but as shown in FIG. 5, for example, after traveling straight for a predetermined distance, the PCR flow path 34 is folded back so as to make a U-turn, and then again in the opposite direction. After going straight for a distance, it may be turned back to make a U-turn, and this may be repeated. At this time, the number of U-turns may be set to any number. The end of such a PCR flow path 34 joins the wash flow path 32.

In the present invention, instead of containing a bactericidal component, a primer for gene detection, and a polymerase enzyme in the liquid to be injected from the sample injection port 33, these substances are sprayed in advance on the inner wall of the PCR flow path 34 or the like. It may be applied. As a result, it is possible to omit the inclusion of the bactericidal component, the gene detection primer, and the polymerase enzyme in the liquid to be injected from the sample injection port 33, and the liquid may be composed of water or the like. Further, when a solvent composed of such a liquid composed of water is injected from the sample injection port 33, the bactericidal component, the gene detection primer, and the polymerase enzyme applied on the inner wall of the PCR flow path 34 are dissolved. Therefore, the desired amplification of nucleic acid fragments can be realized. In such a case, it is desirable that the structure is such that turbulence is generated in the PCR flow path 34 and the mixture can be stirred. Further, it is omitted to include the bactericidal component, the gene detection primer, and the polymerase enzyme in the liquid to be injected from the sample injection port 33, and these bactericidal components, the gene detection primer, and the polymerase enzyme are individually added from the sample injection port 33. It may be injected.

Further, when the substance of Table 1 or Table 2 is previously applied to the surface of the PCR flow path 34 so that the PCR flow path 34 causes turbulent flow and can be stirred, the solution L is used. And the substance applied to the surface of the flow path may be dissolved and mixed. This makes it possible to perform a process for detecting RNA by DNA amplification or cDNA synthesis 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 orthogonal to the straight-ahead direction of the PCR flow path 34. The heating wire 35 is set so that the heating temperature thereof is 90 to 95 ° C. However, this heating temperature is not limited to this, and may be between 30 and 150 ° C. The heating wire 35 is not limited to the case where it is configured in one row, and may be configured in a plurality of rows. In the example of FIG. 5, the width of the heating wire 35 is 5 mm, and the distance between the heating wires adjacent to each other is 5 mm. The PCR flow path 45 passing over such a heating wire 35 passes in a direction orthogonal to the heating wire 35 provided at a predetermined interval, is then folded back, further passes over each heating wire 35, and is further folded back. Then, it repeats passing on each heating wire 35. As a result, the PCR flow path 34 is heated at 90 to 95 ° C. on the heating wire 35, then cooled until it passes over the next heating wire 35, and the temperature drops to about 55 ° C. again. As the next heating wire 35 approaches, the temperature rises and the temperature rises to 90 to 95 ° C. In the process of flowing through the PCR flow path 34, heating and cooling via such a heating wire 35 will repeatedly occur.

The open / close switch 36 controls the washing liquid flowing from the wash flow path 32 to the hybridization flow path 37 by configuring the wash flow path 32 to open and close freely. As a specific configuration of the open / close switch 36, the wash flow path 32 is made of a resin tube, while the open / close switch 36 is made of a film tube that is elastically deformable only at the position of the open / close switch 36. External pressure may be applied to the tube body. By receiving such an external pressure, the film tube is elastically deformed so as to shrink in diameter, so that the passage of the cleaning liquid from the wash flow path 32 can be suppressed. On the other hand, by preventing the external pressure from being applied to the film tube, the film tube does not have a reduced diameter, and in such a case, the cleaning liquid passes through the open / close switch 36 and the hybridization flow path 37. Will flow to. In order to apply this external pressure, for example, an air pump may be installed at the location where the open / close switch 36 is arranged, and the external pressure may be applied by supplying air to the film tube.

A nucleic acid probe is previously formed on the bottom surface of the hybridization flow path 37. In the hybridization flow path 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 flow path 37, for example, when an analysis is performed based on a sample collected from excrement, the following bacteria (bifidobacterium, lactic acid bacterium, Clostridium perfringens, lean bacterium, Frasilis bacterium, butyricum bacterium, equol production) in the intestinal flora are produced. A nucleic acid probe suitable for identifying a bacterium, Clostridium perfringens, Soldery bacterium, Nucretam bacterium, Bacillus cereus bacterium, Copri bacterium, etc. may be arranged.

The hybridization flow path 37 needs to be made of, for example, a light-transmitting material in order to allow light emitted from the outside to reach and to allow light emission to reach the outside. For example, it is made of a transparent plastic material.

In the waste liquid storage reservoir 38, a waste liquid containing nucleic acid other than the target nucleic acid flows from the hybridization flow path 37. The waste liquid storage reservoir 38 is composed of a water storage tank having a storage space for temporarily storing the waste liquid.

The discharge port 39 is provided to discharge the waste liquid accumulated in the waste liquid storage reservoir 38. The discharge port 39 is normally closed by a cap (not shown), but by opening the lid, 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 a lighting unit 41 and a detection unit 42 connected to the control block 46, respectively, and a chip mounter 43 provided so as to face the lighting unit 41 and the detection unit 42, respectively. 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. The nucleic acid analysis chip 3 is attached to the chip mounter 43 so as to be detachable.

FIG. 7 shows the 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 supply stabilization unit 65, an operation unit 66, and a heating wire control. It is equipped with a unit 67. A detection unit 42 and an illumination unit 41 are connected to the central control unit 61, respectively. An air pump 91 and a sensor 92 are connected to the air pump control unit 64. A heating wire 35 is connected to the heating wire control unit 67.

The illumination unit 41 is an illumination that irradiates the nucleic acid analysis chip 3 attached to the chip mounter 43 with light, and is composed of, for example, an LED (blue LED or the like). The illumination unit 41 is arranged so that the irradiated light directly reaches the hybridization flow path 37 in the nucleic acid analysis chip 3. In the illumination unit 41, the central control unit 61 controls ON / OFF of light emission, emission intensity, emission wavelength, emission time, and the like.

The detection unit 42 is a device for receiving a hybridization signal consisting of light emitted from the nucleic acid analysis chip 3 attached to the chip mounter 43, converting the hybrid signal into an electrical signal, and transmitting the signal to the central control unit 61. be. The detection unit 42 is composed of a solid-state image sensor such as a photomultiplier tube and a CCD image sensor. The detection unit 42 is arranged at a position where light emission from the hybridization flow path 37 in the nucleic acid analysis chip 3 can be directly received. In the detection unit 42, the central control unit 61 controls 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 receive and measure 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 the near infrared region from the hybridization flow path 37 by using, for example, a small spectroscope (C12666MA / C12800MA) manufactured by Hamamatsu Photonics. The molecular vibration may be measured with and the substance may be identified. Further, in order to detect minute light such as an ultraviolet region from the hybridization flow path 37, the detection unit 42 is measured using a photomultiplier tube (example: R11265U-200) of Hamamatsu Photonics. May be good.

The chip mounter 43 is provided with a mounting table on which the nucleic acid analysis chip 3 can be placed, 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 to be able to fix the nucleic acid analysis chip 3 so that the hybridization flow path 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 so as to increase the outer surface area in order to easily dissipate heat generated from the heating wire 35. It is desirable that the heat radiating plate 45 is made of a lightweight and easy-to-process material such as aluminum.

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

The communication unit 62 controls to convert the communication signal transmitted from the central control unit 61 into a wireless signal and transmit this to the mobile terminal 5 via the antenna. Further, the communication unit 62 also plays a role of receiving the wireless signal transmitted from the mobile terminal 5, converting it into an electric signal, and supplying it to the central control unit 61. The communication unit 62 is premised on transmitting and receiving wireless signals to and from the mobile terminal 5, but is not limited to this, and communicates with the public communication network 6 without going through the mobile terminal 5. You may do so. Further, the communication unit 62 is not limited to the case where signals are transmitted and received by wireless communication, and may be substituted for wired communication.

The display panel 63 is composed of, for example, a liquid crystal display panel or the like, 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 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 the touch panel, it may be configured integrally with the display panel 63.

The power supply stabilizing unit 65 controls to stabilize the power supply of the control block 46, the heating wire 35, and the like in the chip measuring device 4.

The central control unit 61 is a central control unit for controlling the entire chip measuring device 4, and includes, for example, a CPU (Central Processing Unit). The central control unit 61 transmits various control commands to each unit according to the user's operation via the operation unit 66. Further, the central control unit 61 controls ON / OFF of light emission, light emission intensity, light emission wavelength, light emission time, etc. with respect to the illumination unit 41. Further, the central control unit 61 controls the detection unit 42 to turn on / off the light reception, the light reception time, and the like. The central control unit 61 sends the light receiving information received by the detection unit 42 from the hybridization flow path 37. The central control unit 61 displays such light receiving information on the display panel 63, or transmits it to the outside via the communication unit 62. The central control unit 61 also controls opening / closing by the opening / closing switch 36. The central control unit 61 executes each of these controls based on a program stored in a ROM or the like (not shown).

The air pump control unit 64 controls 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, for example, the detection information by the sensor 92.

The sensor 92 is composed of, for example, a motion sensor, an infrared sensor, or the like, detects the movement of the user's hand, generates detection information based on the sensor 92, 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 has an air tank that stores air to be sent 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. It can be classified as. The air pump 91 is not limited to the case where it is operated by automatic control, and air may be manually supplied to the wash flow path 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 of analyzing a gene using the gene analysis system 1 to which the present invention is applied will be described.

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

When collecting such a sample, the sample is attached to the tip of a rod-shaped body such as a cotton swab. Next, as shown in FIG. 8, the rod-shaped body 9 to which the sample 95 is attached to the tip portion 94 is attached to the collection cartridge 22 in which a plurality of granular bodies 223 are filled in the storage container 220 and the liquid L is immersed. insert. The tip portion 94 of the rod-shaped body 9 may have a slightly larger diameter like a cotton swab, but the diameter is not limited to this, and all rod-shaped bodies may have the same diameter.

As the rod-shaped body 9 is inserted into the storage container 220 from its tip portion 94, the tip 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 to the lower side. You will be guided naturally. By further pushing the tip portion 94 of the rod-shaped body 9 downward from the insertion port 229, the tip portion 94 is inserted into the insertion port 229 and enters the lower chamber 220b. At this time, the protrusion 96 protruding outward may be formed in advance at the root portion of the rod-shaped body 9 slightly separated from the tip portion 94 on the root side. As a result, the protrusion 96 is locked to the protrusion 222 at the insertion port 229, so that the pushing depth of the rod-shaped body 9 can be controlled to an appropriate position, and the rod-shaped body 9 can be placed in the storage container 220. It is possible to prevent the laminated body 26 from being pushed in too much and reaching the laminated body 26. Further, the rod-shaped body 9 may be formed with creases, cut grooves or the like in advance so that the rod-shaped body 9 can be easily folded in the vicinity of the protrusion 96 and easily separated from the tip portion 94.

In the process of pushing the thickened tip portion 94 of the rod-shaped body 9 into the insertion port 229 by preliminarily urging the insertion port 229 formed by the inner wall 221 toward the direction in which the inner diameter thereof is narrowed. The operation of pushing the insertion port 229 outward and inserting it into the lower chamber 220b is performed. Then, when the thickened tip portion 94 is completely inserted into the lower chamber 220b side, the insertion port 229 once expanded is urged again in the direction in which the inner diameter is narrowed. The root portion of the tip portion 94 of the rod-shaped body 9 is closed with the protruding portion 222 of the insertion port 229. As a result, it is possible to create a state in which the tip portion 94 cannot be directly seen by the user. Therefore, if the collected material 95 is excrement, the user cannot directly see it, which makes it possible to give a clean impression.

Further, when the rod-shaped body 9 is pulled out from the lower chamber 220b, the operation of pulling the rod-shaped body 9 toward the upper chamber 220a while expanding the insertion opening 229 outward in the process of pushing out the thickened tip portion 94 from the insertion opening 229. Will be done. At this time, if the sample 95 is attached to the tip portion 94, the sample 95 may be dropped into the lower chamber 220b by applying the urged inner wall 221 to the sample 95. It will be possible.

Further, since the insertion port 229 formed by the inner wall 221 is preliminarily urged in a direction in which the inner diameter thereof is narrowed, the lower chamber 220b is in a state close to being sealed, so that the liquid L inside is formed. And the granular material 223 can be made to last for a long time.

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

The user may agitate the rod-shaped body 9 in the lower chamber 220b, or may cause the rod-shaped body 9 to vibrate up and down. As a result, the collected material 95 adhering to the tip portion 94 is brought into contact with the granular material 223 several times and rubbed, and by wiping the collected material 95 more effectively, it can be dissolved in the liquid L.

Next, the user withdraws the rod-shaped body 9 including the tip portion 94 after the collection 95 has been removed from the storage container 220. Next, as shown in FIG. 9, the collection cartridge 22 is attached to the cartridge accommodating portion 231 in the pretreatment main body 23. As a result, at the time of mounting the collection cartridge 22 and the pretreatment main body 23, the storage container 220 filled with the granular material 223 and the first filter 234 were just separated by the laminated 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, by making a hole in the film 225, the liquid L is allowed to flow down to the separation portion 233 side in the pretreatment main body 23. Any method may be used for making holes in the film 225. For example, in the chamber R1 separated on the upper side via the first filter 234, a plurality of long needles are installed upward. When the collection cartridge 22 is attached, the long needle pierces the film 225, and the liquid L flows down to the separation portion 233 side of the pretreatment main body 23 from the hole made in the film 225. It becomes. On the other hand, since the granular body 223 cannot pass through the reticulated body 224, it is held by the granular body 223 and does not fall to the separation portion 233.

FIG. 10 shows a state in which the film 225 is passed through the liquid L and is allowed to 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 this first filter 234, serum is separated from the liquid L through the pores formed between the continuous tissues of the membrane. That is, this serum will be accumulated in the chamber R1, and the liquid L from which the serum has been removed will pass through the first filter 234 and flow down into the chamber R2.

At least viruses and bacteria are separated from the liquid L flowing down into the chamber R2 by the second filter 235. Only the liquid L from which the virus and the bacterium 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 of the pretreatment body 23 to the sample injection port 33 of 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 flow path 34.

The liquid L guided to the PCR flow path 34 gradually flows down toward the downstream side of this flow path 34. At this time, the liquid L flows through the PCR flow path 34 in the state of a microfluidic by flowing through the reduced diameter PCR flow path 34. The liquid L causes a polymerase chain reaction in the process of flowing through the PCR flow path 34. Specifically, when the PCR flow path 34 is flowing on the heating wire 35, it is in a state of being heated at 90 to 95 ° C. Therefore, at this stage, the nucleic acid is heat-denatured to form a double-stranded nucleic acid. The hydrogen bond will be cleaved and dissociated into a single-stranded nucleic acid.

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

Next, the temperature begins to rise again as the liquid L containing the single-stranded nucleic acid complementarily bound to such a primer gradually approaches the heating wire 35. As the temperature rises, the primer begins to elongate 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, in which hydrogen bonds of the double-stranded nucleic acid are cleaved and dissociated into the single-stranded nucleic acid. It will be.

As a result of the above-mentioned process repeating in the process of flowing the liquid L through the PCR flow path 34, it becomes possible to flow the nucleic acid fragment downstream while amplifying it.

The liquid L will be guided to the hybridization flow path 37 after leaving the PCR flow path 34. At the stage where the liquid L flows through the hybridization flow path 37, at least the hybridization flow path 37 needs to be reached by the illumination light from the illumination unit 41, and light emission from the hybridization flow path 37 is generated. In such a case, it is necessary that the central control unit 61 in the control block 46 controls in advance so that the detection unit 42 can detect this.

In the hybridization flow path 37, if the target nucleic acid is contained in the liquid L, a stable double chain can be formed between the target nucleic acid and the nucleic acid probe. As a result, only the target nucleic acid forms a double chain complementary to this nucleic acid probe and remains in this hybridization flow path 37 as it is, and all nucleic acids other than the target nucleic acid remain in this hybridization flow path 37 as they are. It will flow and be sent to the waste liquid storage reservoir 38.

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

In this way, hybridization is performed in the hybridization flow path 37. In order to more preferably perform hybridization in this hybridization flow path 37, it is necessary that the temperature, pH, cation concentration and the like are prepared in advance. The temperature may be adjusted by using a Perche element, a heating wire, or the like. The temperature may be, for example, about 37 ° C., and the pH may be about 7.

After the hybridization is completed, the washing liquid may be flowed from the wash flow path 32 to the hybridization flow path 37 by opening the open / close switch 36. This makes it possible to identify the nucleic acid multiple times using the nucleic acid analysis chip 3. When the cleaning liquid is flown 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 flowed. The washing liquid flows downstream through the wash flow path 32 and directly flows into the hybridization flow path 37. The washing liquid that has reached the hybridization flow path 37 flows all the remaining nucleic acid and sends it to the waste liquid storage reservoir 38.

When the detection unit 42 can detect the luminescence indicating the presence of the target nucleic acid, the detection unit 42 outputs this to the central control unit 61. After acquiring the detection result, the central control unit 61 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 above data via the communication unit 62, the mobile terminal 5 uses the examination application stored in the mobile terminal 5 to analyze the data to identify the nucleic acid and perform gene analysis. .. The form terminal 5 displays the analysis result to the user, or displays various proposals for maintaining health in the future to the user. Further, the mobile terminal 5 may further transmit the above-mentioned data received via the communication unit 62 to the public communication network 6. The above 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, the knowledge obtained by the information analysis by the central control device 8 and the analysis result may be transmitted to the mobile terminal 5 via the public communication network 6.

According to the present invention having the above-described configuration, the user collects a sample from any of the human body, excrement, and soil with a rod-shaped body 9, and collects the sample on the collection cartridge 22 in which the liquid L is immersed, and on the tip portion 94. The rod-shaped body 9 to which the object 95 is attached is inserted. Other than that, the work of attaching the collection cartridge 22 to the pretreatment main body 23 and the tip of the discharge port 236 in the pretreatment main body 23 are simply connected to the sample injection port 33 of the nucleic acid analysis chip 3 afterwards. All of them will be automatically analyzed.

Therefore, even a user who does not have specialized knowledge about medical treatment can perform gene analysis by himself / herself with an extremely simple task.

Further, since the pretreatment kit 2 for gene analysis, the chip 3 for nucleic acid analysis, and the chip measuring device 4 are not complicated in device configuration, they can be miniaturized and are suitable for self-examination at home. Since it has a structure and can be provided at a low price, it can be purchased by individuals.

Further, according to the present invention, nucleic acids can be extracted from the collected samples with high efficiency, and serum, bacteria, and viruses are removed in advance through the pretreatment kit 2 for gene analysis in order to identify the nucleic acids with high accuracy. Moreover, these operations can be performed extremely easily without the need for special skill or knowledge.

Further, the washing liquid can be washed by flowing the washing liquid from the wash flow path 32 to the hybridization flow path 37, and the nucleic acid analysis chip 3 can be used repeatedly many times, so that the cost performance is very excellent. It has a structure, and this cleaning work can be performed extremely easily.

In addition, while improving the device configuration, ease of handling, and cost performance as described above, the accuracy of biochemical tests can be maintained at the same level as before, which is the same level at medical institutions. It will be possible to realize the gene analysis of the above at home.

The present invention is not limited to the above-described embodiment. The granular body 223 may be configured by pre-sealing the liquid L in the epidermis made of an elastic substance. In such a case, it is not essential to impregnate the liquid L in the storage container 220. When the rod-shaped body 9 to which the collected material is attached to the tip is inserted, the collected material is collected by bringing the tip into contact with the granular body 223 as described above. In addition to this, the tip of the rod-shaped body 9 breaks the epidermis of the granular body 223 to allow the liquid L to flow out. This makes it possible to impregnate the tip of the rod-shaped body 9 with the liquid L that has flowed out.

Further, the gene analysis system 1 to which the present invention is applied may be configured as a case-accommodating type as shown in FIGS. 11 and 12. Since the components and members of the case-accommodating gene analysis system 1'are the same as those in the above-described embodiment, they will be described with the same reference numerals and the following description will be omitted.

FIG. 11 shows the appearance of the case-contained 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 with the hinge (not shown) as the center of rotation. A handle 101 is provided on the upper surfaces of the cases 100a and 100b, and a lock mechanism 102 is provided. Further, a display 103 and a touch panel 104 may be provided on the outside of 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 expanded. The above-mentioned chip measuring device 4 is arranged on the case 100a side, 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 is operated by the electric power supplied from the battery 105. The measurement result by the chip measuring device 4 is sent to this PC 50. The PC 50 has the same function as the mobile terminal 5 described above.

Further, a nucleic acid analysis chip 3 is arranged on the case 100b side, and a pretreatment main body 23 is further connected to the nucleic acid analysis chip 3. The nucleic acid analysis chip 3 is provided at a position facing the chip measuring device 4 when the cases 100a and 100b are closed. This enables the above-mentioned measurement. The discharge port 236 of the pretreatment main body 23 is directly connected to the sample injection port 33 of the nucleic acid analysis chip 3, and 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 is capable of introducing 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 main body 23 via the branch tube 111. Incidentally, the power for sending the liquid L from the collection cartridge 22 to the branch tube 111 is assumed to be performed via the pump 109. The insides of the cases 100a and 100b are covered with a reinforced plastic or a reinforced protective film except for the sample introduction port 108, so that the sample can be prevented from scattering.

According to such a case-accommodating gene analysis system 1', the system can be compactly integrated into a case having a size such as a suitcase. In addition, high-risk viruses and diseases (HIV, Marburg virus, Ebola virus, smallpox, Lassa fever, etc.) can be measured quickly without outsourcing to a special management facility each time, making it a major infectious disease worldwide. It is possible to detect the epidemic as soon as possible and suppress the spread of damage. In particular, according to this case-accommodating gene analysis system 1', the rod-shaped body 9 to which the sample 95 is attached can be simply inserted into the sample introduction port 108, and the rest can be measured fully automatically, so that operability is operability. It is also excellent in terms of hygiene because it can be disposed of without touching any human hands from the start to the end of measurement.

Further, according to the present invention, in addition to nucleic acid measurement, biological substances such as peptides, exosomes, biomarkers, and antibodies can be measured in the same manner. In such a case, the liquid L injected through the discharge port 236 in the pretreatment main body 23 will be directly flowed to the hybridization flow path 37 without nucleic acid amplification in the PCR flow path 34. All other steps are the same as those of 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, a injection flow path for simply flowing the injected liquid L into the hybridization flow path 37. It suffices if it has been done.

1 Gene analysis system 2 Pretreatment kit for gene analysis 3 Chip for nucleic acid analysis 4 Chip measuring device 5 Mobile terminal 6 Public communication network 7 Server 8 Central control device 9 Rod 12 Public communication network 21 Plunger 22 Collection cartridge 23 Pretreatment Main body 26 Laminated body 31 Air pump connection port 32 Wash flow path 33 Sample injection port 34 PCR flow path 35 Heating wire 36 Open / close switch 37 Hybridization flow path 38 Waste liquid storage reservoir 39 Discharge port 41 Lighting 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 Collected material 96 Projection part 211 Inner cylinder part 212 Pusher 213 Rubber part 220 Storage container 220a Upper room 220b Lower room 221 Inner wall 222 Protruding part 223 Granules 224 Reticulated body 225 Film 229 Insertion port 231 Cartridge storage part 232 Peripheral wall 233 Separation part 234 First filter 235 Second filter 236 Discharge Exit 237 cap

Claims (7)

In a pretreatment kit for gene analysis for extracting biological substances from collections collected from the human body, excrement, or soil.
The epidermis is filled with a plurality of granules composed of an elastic substance, and a liquid containing a bactericidal component, a primer for gene detection, and a polymerase enzyme is enclosed in the epidermis, and the collected material is attached to the tip. When a rod-shaped body is inserted, the sample is collected by bringing the tip into contact with the granules, and the collection cartridge is impregnated with the liquid that has flowed out by breaking the epidermis with the tip. ,
A discharge means for discharging the liquid sent from the collection cartridge to the outside ,
A first filter for separating at least serum from the liquid delivered from the mounted collection cartridge, and a pretreatment body having the discharge means are provided.
The discharging means discharges the liquid that has passed through the first filter to the outside, and discharges the liquid to the outside.
When the collection cartridge and the pretreatment main body are attached, the storage container filled with the granules and the first filter are a reticulated body through which the liquid can pass and a film through which the liquid cannot pass. Are separated by a laminate that is laminated to each other,
When the liquid is delivered from the collection cartridge, the film is formed with holes for passing the liquid so that the liquid can pass through the film, and the granules are held by the network. Pretreatment kit for use. The pretreatment kit for gene analysis according to claim 1 and
A nucleic acid analysis chip for analyzing a target nucleic acid to be detected from a collection collected from the human body, excrement, or soil, and a gene detection primer, a polymerase enzyme, and a biological substance extracted from the above collection. This is hybridized with a PCR channel in which the contained liquid is injected and the nucleic acid fragment in the target nucleic acid is amplified and flows downstream, and an amplification product that is continuous from the PCR channel and flows in from the PCR channel. A nucleic acid analysis chip having a hybridization flow path flowing downstream and a target nucleic acid analysis means for identifying the target nucleic acid by measuring a hybridization signal generated in the hybridization flow path is provided.
An analysis system characterized in that the liquid discharged from the discharge port in the pretreatment kit for gene analysis flows into the PCR flow path in the chip for nucleic acid analysis. It also has at least a wash channel for the cleaning liquid to flow.
The second aspect of the present invention is characterized in that the hybridization flow path is continuous from the wash flow path and can be washed by flowing in the washing liquid from the wash flow path after the hybridization. Analysis system. The PCR flow path is configured to have a diameter on the order of micron, and is arranged so as to pass on a heating wire having a heating temperature of 30 to 150 ° C. provided at predetermined intervals, and the target when passing on the heating wire. The analysis system according to claim 2 or 3, wherein the nucleic acid is thermally denatured. The PCR flow path is arranged so as to repeatedly pass on the heating wire provided at a predetermined interval, be folded back, pass on the heating wire, and then be folded back and pass on the heating wire. 4. The analysis system according to claim 4. The analysis system according to claim 3, wherein the wash flow path alternately flows a cleaning liquid and air. The analysis system according to any one of claims 2 to 6, further comprising a communication means for transmitting information detected by the target nucleic acid analysis means to the outside via a communication network.

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