JP6539935B2 - Separation filter, pretreatment kit for gene analysis - Google Patents

Description

  The present invention relates to a separation filter for extracting a biological substance from a collected material collected from any of human body, excrement and soil, a chip for nucleic acid analysis for analyzing a target nucleic acid to be detected from the collected material, and gene analysis It is about the system.

  Conventionally, medical examinations have been conducted exclusively at specialized medical institutions such as hospitals, but in Japan, with the arrival of a super-aging society, the burden of examinations at medical institutions has become a serious situation. For this reason, while an examination that requires a dedicated device such as X-ray imaging and measurement of an electrocardiogram, and an examination requiring a syringe such as a blood test etc. continue to be conducted at a medical institution, the user simply performs an examination. What can be done should also be considered to reduce the burden of medical checkup at medical institutions by using self-examination conducted at home. If self-examination to be conducted at home can be established, therapeutic medicine and preventive medicine can be enhanced, for example, even in developing countries where necessary medical services are not sufficiently provided.

  Under these circumstances, in order to enable next-generation self-examination, it is possible to realize even smaller users of testing equipment, generalization, lower prices, and even users who do not have specialized knowledge about medical care. A degree of ease of handling is required.

  In response to such social needs, genetic analysis using a chip for nucleic acid analysis (DNA chip) has been realized in recent years. The chip for nucleic acid analysis is used in various fields such as medical field, chemical field, drug discovery field, clinical examination field, food field, agricultural field, engineering field, forensic field and criminal identification field, for example, medical field In general, examinations of various diseases including cancer are generally performed by examination using biochemical samples such as blood.

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

  Subsequently, the presence or absence and / or amount of the target nucleic acid is detected by scanning the hybridization signal from the nucleic acid analysis chip after washing. In this scanning, laser light of a wavelength suitable for exciting a fluorescent substance is irradiated and scanned on the substrate of the chip for nucleic acid analysis. Thereby, the amount of luminescence of the nucleic acid bound to each spotted nucleic acid probe is measured, and based on that, it becomes possible to carry out the analysis process.

  For the purpose of meeting the needs for self-examination conducted at home, in the chip for nucleic acid analysis as described above, for example, the technology shown in Patent Document 1 is disclosed. This Patent Document 1 discloses a nucleic acid analysis chip which can detect a reaction instantaneously, has a small and portable analyzer configuration, stores reaction reagents in advance, and can be disposed together with the reagents 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.

Unexamined-Japanese-Patent No. 2004-28589 JP-A-2014-529083

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

  Patent Document 1 proposes that a chip for nucleic acid analysis is provided with an adsorption groove, and the chip is adsorbed and fixed at a desired position by vacuum suction via a suction groove by a vacuum evacuation pump. However, when such a configuration is adopted, it becomes a large barrier to miniaturizing the analyzer.

  Accordingly, the present invention has been made in view of the above-mentioned problems, and the object of the present invention is to miniaturize and generalize an inspection device in a nucleic acid analysis chip that enables next-generation self-examination. In addition to price reduction, ease of handling that can be achieved by users without specialized knowledge about medical treatment is ensured, and analysis with a nucleic acid analysis chip that can further improve the accuracy of biochemical examination It is an object of the present invention to provide a pretreatment kit for gene analysis for extracting a biological substance from a collected product collected from any of a human body, excrement and soil in a pre-processing stage, and a separation filter used therefor.

The separation filter according to the first aspect of the present invention is a separation filter for separating at least serum or plasma from a liquid containing a collected material from any of human body, excrement and soil, comprising a membrane filter and the membrane filter. The separation filter is formed of either a hollow fiber or a porous material, and is formed of two or more layers, and is laminated on a local region on the membrane filter in a plan view. The boundary line in plan view with respect to the membrane filter is characterized in that at least the upper end is arc-shaped in side view .

  A separation filter according to a second aspect of the present invention is the separation filter according to the first aspect, wherein the separation filter is composed of two or more layers, and is laminated on a local region on the membrane filter in a plan view. The boundary line of the present invention is characterized in that at least an end portion in a plan view has an arc shape or a substantially semicircular shape.

  The separation filter according to a third aspect of the present invention is characterized in that, in the first aspect or the second aspect, the anti-coagulation agent is added with an anticoagulant or a biological substance.

The pretreatment kit for gene analysis according to the fourth aspect of the present invention is a pretreatment kit for gene analysis for extracting a biological substance from a collected product collected from any of a human body, excrement, and soil, in which a plurality of particles are filled. Liquid is immersed in the granular material, and when a rod-like body having the collected matter attached to the tip is inserted, the collected matter is collected by bringing the tip into contact with the granular material, and At least plasma or serum from the collection cartridge to be impregnated with the liquid, the discharging means for discharging the liquid supplied from the collection cartridge to the outside, and the liquid supplied from the mounted collection cartridge They were separated, and the membrane filter, is laminated on the membrane filter, hollow fibers, and a separating filter which is composed of either a porous material A separation filter for, further comprising a processing body before having the discharge means, said discharge means, characterized by discharging the liquid that has passed through the separation filter to the outside.

  In the pretreatment kit for gene analysis according to the fifth invention, in the fourth invention, the body for pretreatment further has a second filter for separating at least virus and bacteria from the liquid which has passed through the separation filter. The discharge port discharges the liquid having passed through the second filter to the outside.

  According to the present invention having the above-described configuration, the user collects the collected material from any of the human body, excrement, and soil with the rod-like body, and the collected material is attached to the tip of the collection cartridge in which the liquid is immersed. Insert the rod-like body. In other cases, simply attach the collection cartridge to the pretreatment body and simply connect the tip of the discharge port of the pretreatment body to the sample inlet of the chip for nucleic acid analysis, and then all the genes will be automatically generated. Analysis will be performed.

  For this reason, even a user who does not have specialized knowledge about medical care can perform genetic analysis by himself with extremely simple work.

  In addition, since the pretreatment kit for gene analysis, the chip for nucleic acid analysis, and the chip measurement device are all not complicated in apparatus configuration, miniaturization is possible, and a structure suitable for self-examination at home It can be purchased by individuals because it can be provided inexpensively.

  Furthermore, according to the pretreatment kit for gene analysis of the present invention and the serum separation filter applied thereto, it is possible to separate plasma or serum with high efficiency.

It is a figure showing the system configuration of the gene analysis system to which the present invention is applied. It is a figure for demonstrating the structure of the cartridge for collection. It is a figure for demonstrating the detail of the laminated body comprised by two-layer structure of a mesh 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 measurement 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 the cartridge for collection | collection in the cartridge accommodating part in the main body for pre-processing. It is a figure which shows the state which allowed the film to pass through per liquid, and was made to flow down to the separation part side in the pre-processing main body. 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. It is a figure which shows the example of a form of the filter for isolate | separating plasma or serum. FIG. 6 is a diagram for explaining an example of use of a filter for separating plasma or serum. It is a figure which shows the other embodiment of the filter for isolate | separating plasma or serum.

  Hereinafter, an embodiment for implementing 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 a pretreatment kit 2 for gene analysis, a chip for nucleic acid analysis (DNA chip) 3, a chip measurement apparatus 4 on which the chip 3 for nucleic acid analysis is mounted, and the chip measurement apparatus 4 The portable terminal 5, a server 7 connected to the portable terminal 5 via the public communication network 6, and a central control unit 8 connected to the public communication network 6 are provided.

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

  The portable 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 portable terminal 5 to confirm his own examination record or confirm various proposals for health maintenance in the future. 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 in which the portable terminal 5, the server 7 and the central control unit 8 are connected via a communication line. The public communication network 6 may be configured as 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 in the server 7. In this database, the information sent via the public communication network 6 is accumulated. Further, the server 7 transmits the stored information to the portable terminal 5 via the public communication network 6 based on a request from the portable terminal 5.

  The central control unit 8 is constituted by 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, this central control device 8 may be operated by a specialist such as a doctor, or the information stored in the server 7 may be used. The knowledge obtained by analyzing information by the central control unit 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 unit 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 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 cylindrical portion 211 and a pusher 212. The inner cylindrical portion 211 is extended in the longitudinal direction, and a rubber portion 213 is provided at the extended end. 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 cylindrical portion 211 is collected by a cartridge by pressing and pressing a user's finger. 22 is configured to be insertable.

  As shown in FIG. 2, the collecting cartridge 22 is filled with a plurality of granular bodies 223 in a 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 which bulges inward and a laminate 26 provided on the bottom of the storage container 220.

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

  The granular body 223 will be described by way of example in the case of being constituted by spherical beads, but it is not limited to this and may be constituted by any polyhedron including cubic and rectangular solid. , And may be configured in a random three-dimensional shape. Moreover, this granular material 223 may be comprised, for example in a specific shape like a hook shape.

  The surface of the granular body 223 may be further coated with calcium phosphate. It is possible to cause the pH of the storage container 220 to approach neutrality through this calcium phosphate layer. Further, the surface of the granular body 223 is coated with, for example, any of zeolite, an emulsion gel, a hydrophilic polymer, and an amino acid substance so that feces and solid as excrement are adsorbed to the granular body 223, It becomes possible to make it easy to take extra things with a filter.

  The liquid L is previously 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 collected material to be extracted from genomic DNA as a biological substance is a 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 is expressed 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 comprises HCl, EDTA, NaCl, SDS and 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 DNA degradation inhibition. DNase / RNase-treated water is water from which DNase, which is a DNA degrading enzyme, is removed, and is composed of a DNA degrading enzyme and water from which an RNA degrading enzyme has been removed.

  If the target product is blood or urine, it may be composed of the composition 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 adjustment buffer is a stabilizing substance for preventing blood coagulation and comprises EDTA, SDS or the like. The buffer solution is a solution for preventing degradation 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, which is a DNA degrading enzyme, is removed, and is composed of a DNA degrading enzyme and water from which an RNA degrading enzyme has been removed.

  If the subject matter to be collected 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 degradation and inactivation of genomic DNA, and is formed by mixing citric acid and disodium hydrogen phosphate. The DNase / RNase-treated water is water from which DNase, which is a DNA degrading enzyme, is removed, and is composed of a DNA degrading enzyme and water from which an RNA degrading enzyme has been removed. The pronase solution is a solution for removing a protein called mucin contained in saliva, and consists of lactose, hydroxypropyl cellulose, pronase and the like.

  The inner wall 221 is made of an elastic material including rubber and the like, and is configured so as to be elastically expanded in diameter and reduced in diameter in the radial direction of the storage container 220. The inner diameter of the inner wall 221 is gradually reduced in diameter from the upper end of the storage container 220 downward, and the inner diameter is the smallest at the middle stage. In the middle stage where the inner diameter is the smallest, an insertion port 229 surrounded by a projecting portion 222 further projecting inward is provided. The inner diameter of the insertion port 229 is formed so as to gradually increase from the insertion opening 229 downward. As a result, the storage container 220 is divided into two chambers, the upper chamber 220a and the lower chamber 220b, bordering the insertion port 229 via the inner wall 221.

  The inner cylindrical portion 211 of the above-described plunger 21 is insertable into the upper chamber 220a. Granules 223 are filled in the lower chamber 220b, and the liquid L is further injected. The particulates 223 and the liquid L are basically charged into 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 biased 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 be bulged inward. That is, all the inner diameters of the inner wall 221 may be the same without changing.

  The laminated body 26 is comprised by the 2 layer structure of the reticulated body 224 and the film 225, as shown in FIG. 3, and these are mutually bonded and comprised.

  The mesh body 224 is formed of, for example, a mesh structure or a mesh in which warp and weft yarns are woven, and the mesh of the mesh has a diameter through which the liquid L can pass. However, the mesh-like body 224 is required to have a diameter such that the mesh 223 can not pass the granular material 223 described above. As a result, even if the liquid L passes through the mesh body 224, the passage of the granular material 223 can be prevented, and the granular material 223 can be held.

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

  The mesh body 224 and the film 225 may be laminated by being bonded to each other, or may be integrated and laminated. Also, only the circumferential end portions of the mesh body 224 and the film 225 may be attached to each other. Further, the mesh body 224 and the film 225 are not limited to being 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 220 b in the storage container 220. As a result, the stacked body 26 plays a role as the bottom of the storage container 220. The plane configuration of the reticulated body 224 and the film 225 constituting the laminate 26 is in accordance with the plan view of the lower chamber 220b to which it is attached.

  As shown in FIG. 4, the pre-processing main body 23 has a cartridge accommodating portion 231 and a separating portion 233 extended downward from the cartridge accommodating 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 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, for example, glass, resin, metal, and ceramics.

  The cartridge housing portion 231 is configured such that the periphery thereof is surrounded by the peripheral wall 232, and the collection cartridge 22 is inserted and configured to be mountable.

  Although the separation portion 233 is configured to gradually decrease in inner diameter toward the lower side, the present invention is not limited to this, and all the inner diameters may be the same. The separation portion 233 is in the form of a tube which can be guided from the cartridge storage portion 231 continuously 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 a distance from the first filter 234 toward the lower side.

  The first filter 234 is composed of a membrane capable of separating at least plasma or serum from the liquid L. That is, the plasma or serum separated from the liquid L is retained in the chamber R1 in the chamber R1 separated on the upper side via the first filter 234 and the chamber R2 separated on the lower side, and the plasma or serum It plays a role of transmitting the separated liquid L to the chamber R2.

  As the first filter 234, "Microza MF" manufactured by Asahi Kasei Chemicals Corporation, "silicone hollow fiber" manufactured by Fuji Systems Co., Ltd., "Zalto clear depth filter 290-C4" manufactured by Sarutrius Stedum Biotech, etc. may be used. May be The first filter 234 is composed of a hollow fiber membrane and separates plasma or serum from the liquid L through the pores formed between the continuous tissues of the membrane.

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

  As the second filter 235, “Microza UF” manufactured by Asahi Kasei Chemicals Co., Ltd., “Zalto 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 virus and bacteria are separated is delivered. The discharge port 236 is formed at the tip of a tubular shape projecting downward, so that the convenience at the time of injecting the liquid L toward the nucleic acid analysis chip 3 can be improved.

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

  Incidentally, this nucleic acid analysis chip 3 is not limited to the case where the collection cartridge 22 and the pretreatment main body 23 are separated from each other, and they are integrated with each other. It is also good. In the pre-processing main unit, the configurations of the first filter 234 and the second filter 235 may be omitted, and in such a case, only the means for discharging the liquid from the collection cartridge 22 is mounted. Just do it. Specifically, this is 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 inlet 33, and the sample inlet 33. The PCR flow channel 34, the heating wire 35 provided in proximity to the PCR flow channel 34, the open / close switch 36 provided at the downstream end of the wash flow channel 32, and the wash flow channel 32 and the PCR flow channel 34 A hybridization channel 37 provided on the further downstream side of the combined downstream, a waste solution storage reservoir 38 provided on the downstream side of the hybridization channel 37, and a discharge port 39 continuing to the waste solution reservoir 38 Is equipped.

  An air pump (not shown) is connected to the air pump connection port 31 to feed air. The air and the cleaning solution fed 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 channel 32 is a flow channel for alternately flowing the air and the cleaning liquid. In order to flow such air and cleaning solution alternately, a bifurcated air tube is attached to the inlet of the wash flow passage 32. Then, the cleaning solution flows from one of the branched tubes, and air is sent from the tube to the branched one via a micro pump. By increasing the pressure of the air sent from the micro pump, the air enters between the cleaning solutions, and the air and the cleaning solutions flow alternately. This can also be realized by periodically fluctuating the amount of air or pressure sent from the micro pump.

  The interval between the alternately flowing air and the cleaning solution may be arbitrary. The wash flow channel 32 is not limited to the case where the air and the cleaning liquid flow alternately. That is, the wash flow channel 32 may flow only the cleaning liquid. The wash flow channel 32 may be omitted.

  The sample inlet 33 is connectable to the tip of the outlet 236 in the pretreatment main body 23 described above. The liquid L is injected into the sample inlet 33 through the connected outlet 236. The sample inlet 33 may be closed by a lid not shown when not in use. The sample inlet 33 is not necessarily limited to the case where the tip of the outlet 236 in the pretreatment body 23 is connectable. In such a case, the discharge port 236 in the pretreatment main body 23 may be positioned immediately above the sample inlet 33 in a state of being separated from the sample inlet 33, and the liquid L may be dropped to the sample inlet 33. That is, the sample inlet 33 may have any configuration as long as the liquid L can be fed from the outlet 236 of the pretreatment body 23. The liquid L fed into the sample inlet 33 is led 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 cross-sectional form of the PCR channel 34 may be any form such as circular and polygonal, but it is desirable that its diameter be in the micron order of less than 1 mm. As a result, the liquid L passing through the PCR flow channel 34 is in a so-called micro fluid state, and conversely, the PCR flow channel 45 for causing such micro fluid to flow down is in a so-called micro flow channel. In addition, in order to allow the liquid L to efficiently flow downstream in the state of microfluidics, the height may be set to gradually decrease as the PCR flow channel 34 proceeds downstream.

  The PCR channel 34 may be any path, but, for example, as shown in FIG. 5, it is folded to make a U-turn after going straight for a predetermined distance, and then it is turned again in the opposite direction. After going straight through the distance, it may be folded 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 channel 34 joins the wash flow channel 32.

  In the present invention, instead of containing the bactericidal component, the primer for gene detection, and the polymerase enzyme in the liquid to be injected from the sample injection port 33, these substances are previously sprayed or the like on the inner wall of the PCR channel. 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 made of water or the like. In addition, when a solvent composed of a liquid composed of such water is injected from the sample inlet 33, the bactericidal component, the gene detection primer, and the polymerase enzyme applied on the inner wall of the PCR channel 34 are dissolved. Because it is possible to achieve the desired amplification of the nucleic acid fragment. In such a case, it is desirable that turbulent flow occurs in the PCR flow channel 34 to make it possible to stir. In addition, the bactericidal component, the primer for gene detection, and the polymerase enzyme are not included in the liquid to be injected from the sample inlet 33, and the bactericidal component, the primer for gene detection, and the polymerase enzyme are separately provided from the sample inlet 33. It may be injected.

  In addition, when the substances in Table 1 or Table 2 are applied in advance on the surface of the PCR flow channel 34, and the PCR flow channel 34 has a structure that causes turbulent flow and can be stirred, the solution L The substance applied to the surface of the flow path may be dissolved and mixed. This makes it possible to perform processing for detecting RNA by DNA amplification or cDNA synthesis in the PCR channel 34.

  The heating wire 35 is provided below the PCR flow channel 34. The heating wire 35 is extended so that the longitudinal direction thereof is orthogonal to the direction in which the PCR flow path 34 goes straight. The heating wire 35 is set to have a heating temperature of 90 to 95 ° C. However, this heating temperature is not limited to this, and it may be between 30 and 150 ° C. The heating wires 35 are not limited to being 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 adjacent heating wires is 5 mm. The PCR flow path 45 passing above the heating wire 35 passes in the direction orthogonal to the heating wire 35 provided at a predetermined interval, and then is folded back, passes further above the heating wire 35, and is further folded back. Thus, passing over each heating wire 35 is repeated. As a result, the PCR channel 34 is heated at 90 to 95 ° C. on the heating wire 35 and then cooled before passing on 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, such heating and cooling via the heating wire 35 will occur repeatedly.

  The open / close switch 36 controls the washing solution flowing from the wash flow channel 32 to the hybridization flow channel 37 by configuring the wash flow channel 32 so as to be able to open and close. As a specific configuration of the open / close switch 36, while the wash flow passage 32 is formed of a resin tube, it is formed of a film tube which is elastically deformable only at the position of the open / close switch 36. An external pressure may be applied to the tube. By being elastically deformed so as to reduce the diameter of the film tube by receiving such an external pressure, it is possible to suppress the passage of the cleaning liquid from the wash flow channel 32. On the other hand, by not applying an external pressure to the film tube, the diameter of the film tube is not reduced, and in this case, the washing solution passes through the open / close switch 36 and the hybridization channel 37 It will flow to the In order to apply the external pressure, for example, an air pump may be installed at a position where the open / close switch 36 is disposed, and the external pressure may be loaded by supplying air to the film tube.

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

  In the hybridization channel 37, for example, when analysis is performed on the basis of a certain collected product from excrement, the following bacteria in the intestinal bacterial flora (Bifidobacterium, Lactobacillus, Ficus. A nucleic acid probe suitable for identifying bacteria, Welsh bacteria, Solderi bacteria, Nucletam bacteria, Bacillus cereus, copuri bacteria etc.) may be arranged.

  The hybridization channel 37 needs to be made of, for example, a light transmitting material in order to allow the light irradiated from the outside to reach as well as the light emission to reach the outside. For example, it is made of a transparent plastic material.

  In the waste fluid storage reservoir 38, waste fluid containing nucleic acids other than the target nucleic acid flows from the hybridization channel 37. The waste solution storage reservoir 38 is constituted by a water storage tank comprising a storage space capable of temporarily storing the waste solution.

  The discharge port 39 is provided to discharge the waste liquid accumulated in the waste liquid storage reservoir 38. Although the discharge port 39 is normally closed by a cap (not shown), by opening the lid, it is possible to discharge the waste liquid accumulated in the waste liquid storage reservoir 38.

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

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

  FIG. 7 shows a block configuration of this 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 generation stabilizing unit 65, an operation unit 66, and heating wire control. And a unit 67. The central control unit 61 is connected to the detection unit 42 and the illumination unit 41 respectively. An air pump 91 and a sensor 92 are connected to the air pump control unit 64. The heating wire control unit 67 is connected to the heating wire 35.

  The illumination unit 41 is an illumination that emits light to the nucleic acid analysis chip 3 attached to the chip mounter 43, and is configured of, for example, an LED (blue LED or the like). The illumination unit 41 is disposed 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 hybridization signal consisting of light emission from the nucleic acid analysis chip 3 attached to the chip mounter 43, converting this into an electrical signal, and transmitting it to the central control unit 61. is there. The detection unit 42 is formed of, for example, a solid-state image sensor such as a photomultiplier tube or a CCD image sensor. The detection unit 42 is disposed at a position where light emission from the hybridization channel 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 the case of 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, this detection unit 42 absorbs light in the infrared region and near-infrared region from the hybridization flow channel 37 using, for example, a small spectroscope (C12666MA / C12800MA) manufactured by Hamamatsu Photonics, etc. The molecular vibration may be measured by the above to identify the substance. In addition, in order to detect minute light in the ultraviolet region and the like from the hybridization flow channel 37, measurement is performed using a photomultiplier tube (eg, R11265U-200) of Hamamatsu Photonics for this detection unit 42. It is 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 as to be able to detachably fix the mounting table. The chip mounter 43 is configured to be able to fix the nucleic acid analysis chip 3 so that the hybridization channel 37 is oriented in the direction of the illumination unit 41 and the detection unit 42.

  The heat dissipation plate 45 is provided in the vicinity of the heating wire 35, and in order to easily dissipate the heat generated from the heating wire 35, the heat dissipation plate 45 has a shape provided with unevenness so as to increase the surface area of the outside. It is desirable that the heat sink 45 be made of a lightweight material such as aluminum, which is easy to process.

  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 converts a communication signal sent from the central control unit 61 into a wireless signal, and controls transmission of the signal to the portable terminal 5 via an antenna. The communication unit 62 also plays a role of receiving a radio signal transmitted from the portable terminal 5, converting the signal into an electric signal and supplying the signal to the central control unit 61. The communication unit 62 is premised on transmitting and receiving a radio signal to and from the portable terminal 5, but is not limited to this, and communicates with the public communication network 6 without passing through the portable terminal 5. You may do so. Further, the communication unit 62 is not limited to the case of transmitting and receiving a signal by wireless communication, and may be replaced by wired communication.

  The display panel 63 is, 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 is configured of a button, a mouse, a touch panel, and the like for the user to operate and control the chip measurement 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, this may be integrally configured with the display panel 63.

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

  The central control unit 61 is a central control unit for controlling the entire chip measurement device 4 and includes, for example, a CPU (Central Processing Unit). The central control unit 61 transmits various control commands to each unit via the operation unit 66 according to the user's operation. Further, the central control unit 61 controls ON / OFF of light emission, light emission intensity, light emission wavelength, light emission time and the like with respect to the illumination unit 41. Further, the central control unit 61 controls the detection unit 42 to turn on / off light reception, light reception time, and the like. The central control unit 61 receives the light reception information received by the detection unit 42 from the hybridization channel 37. The central control unit 61 displays such light reception information on the display panel 63 or transmits it via the communication unit 62 to the outside. 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 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 detection information by the sensor 92, for example.

  The sensor 92 is configured 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 movement, and transmits it to the air pump control unit 64. As a result, 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 is an air tank for storing air to be supplied by sucking in air from the outside, and a tube for supplying 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 flow path 32 through the air pump connection port 31. In such a case, the configurations 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 collected material collected from any of human body, excrement and soil. The collection target of the collection here is blood, sweat, saliva or the like if it is the human body, and stool or urine if it is excrement.

  When such a collected material is collected, the collected material 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 matter 95 attached to the tip end portion 94 is placed on the collection cartridge 22 in which the plurality of granular bodies 223 are filled in the storage container 220 and the liquid L is immersed. insert. The tip end portion 94 of the rod-like body 9 may be slightly thickened like a cotton swab, but it is not limited to this, and all rod-like bodies of the same diameter may be used.

  As the rod-like body 9 is inserted into the container 220 from its tip 94, the tip of the rod-like body 9 is directed toward the insertion port 229 via the inner diameter of the inner wall 221 gradually reduced in diameter from the upper end downward. It will be guided by nature. When the distal end 94 of the rod 9 is further pressed downward from the insertion opening 229, the distal end 94 is inserted into the insertion opening 229 and enters the lower chamber 220b. At this time, the protruding portion 96 may be formed in advance at the base portion slightly apart from the tip end portion 94 of the rod-like body 9 and protruding outward. As a result, the projection 96 is locked to the projection 222 at the insertion port 229, so that the pushing depth of the rod 9 can be controlled to an appropriate position. It is possible to prevent the sheet 26 from being pushed too much and reaching the stack 26. Furthermore, a fold or a kerf may be formed in advance so that the rod-like body 9 can be easily broken in the vicinity of the projection 96 and easily separated from the tip 94.

  The insertion opening 229 formed by the inner wall 221 is biased in advance in the direction in which the inner diameter is narrowed, thereby pushing the enlarged diameter distal end portion 94 of the rod 9 into the insertion opening 229. While spreading the insertion port 229 outward, an operation to insert this into the lower chamber 220b side is performed. Then, when the diameter-increased end portion 94 completely enters the lower chamber 220 b side, the insertion port 229 once pushed out is biased again in the direction in which the inner diameter narrows, The root portion of the distal end portion 94 of the rod-like body 9 closes the projecting portion 222 of the insertion port 229. As a result, it is possible to create a state in which the user can not directly view the tip 94. For this reason, if the collected item 95 is excrement, it is possible for the user to have a clean impression because the user can not directly view it.

  Also, when pulling out the rod-like body 9 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 pushed outward and is pulled toward the upper chamber 220a. Will do. At this time, if the collected matter 95 is attached to the tip end portion 94, the collected matter 95 may be dropped into the lower chamber 220b by applying the inner wall 221 subjected to the biasing force to the collected matter 95. It becomes possible.

  Furthermore, since the lower chamber 220b is in a state close to sealing because the insertion port 229 formed by the inner wall 221 is biased in the direction in which the inner diameter is narrowed, the liquid L inside And the granular body 223 can be made to last longer.

  The periphery of the tip end portion 94 of the rod-like body 9 which has entered the lower chamber 220 b in this manner is surrounded by the granular body 223. As a result, since the granular material 223 comes in contact with the tip end portion 94, the collected matter 95 adhering to the distal end portion 94 also comes in contact with the granular material 223. As a result, the collected matter 95 is wiped off and collected by the granular body 223 in contact with it. Since the granular material 223 is impregnated in the liquid L, the collected material collected is dissolved in the liquid L.

  The user may stir the rod-like body 9 in the lower chamber 220 b or may slightly shake the rod-like body 9 vertically. As a result, the collected matter 95 attached to the tip portion 94 is brought into contact with the granular material 223 several times to be rubbed, and the collected matter 95 can be dissolved in the liquid L by wiping the collected matter 95 more effectively.

  Next, the user withdraws the rod-like body 9 including the tip 94 from which the collected material 95 has been removed from the storage container 220. Next, as shown in FIG. 9, the collecting cartridge 22 is mounted in the cartridge accommodating portion 231 of the pre-processing 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 are just separated via the stacked body 26. It becomes a state. In this state, the liquid L is held by the film 225 which can not pass through the liquid itself.

  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. Although any method may be used as a method of making a hole in the film 225, for example, in a chamber R1 separated on the upper side via the first filter 234, a long needle is installed upward across a plurality of needles. When the collection cartridge 22 is attached, the long needle is stuck in the film 225, and the liquid L flows down from the hole opened in the film 225 to the separation unit 233 side in the pretreatment main body 23 It becomes. On the other hand, since the granular material 223 can not pass through the reticulated body 224, the granular material 223 does not fall into the separating portion 233 by being held there.

  FIG. 10 shows a state in which the film 225 is allowed to pass through the liquid L and flowed down to the separating 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, the serum is accumulated in the chamber R1, and the liquid L from which the serum is removed passes through the first filter 234 and flows down to the chamber R2.

  At least virus and bacteria are separated by the second filter 235 from the liquid L flowing down into the chamber R2. Only the liquid L from which virus and bacteria have been separated passes through the second filter 235 and flows down to the room R3.

  After accumulating the liquid that has flowed down into the room R3, the user opens the cap 237 and connects the tip of the outlet 236 in the pretreatment main body 23 to the sample inlet 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 of the pretreatment body 23. The liquid L fed into the sample inlet 33 is led to the PCR channel 34.

  The liquid L introduced to the PCR channel 34 will gradually flow downward toward the downstream of the channel 34. At this time, the liquid L flows in the PCR channel 34 in the state of the microfluidic by flowing in the reduced diameter PCR channel 34. The liquid L causes the polymerase chain reaction in the process of flowing through the PCR channel 34. Specifically, when the PCR flow channel 34 is flowing on the heating wire 35, it is in a state of being heated at 90 to 95 ° C. The hydrogen bond is broken and dissociated into single-stranded nucleic acid.

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

  Next, as the liquid L containing single-stranded nucleic acid complementarily bound to such a primer gradually approaches the heating wire 35, the temperature starts to rise again. In response to this temperature rise, the primer begins to extend and becomes a double stranded nucleic acid. Then, the liquid L reaches the heating wire 35 again, and heating occurs at a temperature of 90 to 95 ° C., thereby causing denaturation, breaking the hydrogen bond of the double stranded nucleic acid, and dissociating into single stranded nucleic acid. It will be.

  As the liquid L flows through the PCR channel 34, the above-described process is repeated, so that the nucleic acid fragment can be amplified and flowed downstream.

  The liquid L is led to the hybridization channel 37 after leaving the PCR channel 34. At the stage where the liquid L flows through the hybridization channel 37, at least the hybridization channel 37 needs to have illumination light from the illumination unit 41, and light emission from the hybridization channel 37 occurs. In this case, the central control unit 61 in the control block 46 needs to control in advance a state in which the detection unit 42 can detect this.

  In the hybridization channel 37, if the target nucleic acid is contained in the liquid L, it becomes possible to form a stable duplex between it and the nucleic acid probe. As a result, only the target nucleic acid remains in the hybridization channel 37 as it is by forming a double strand complementary to the nucleic acid probe, and any nucleic acid other than the target nucleic acid remains in the hybridization channel 37 as it is. It flows and is fed to the waste solution 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 measurement device 4. The hybridization performed in the hybridization channel 37 may be performed by any known method. For example, in the case of employing 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. It may be detected by

  In this way, hybridization is performed in the hybridization channel 37. In order to carry out hybridization more suitably in the hybridization channel 37, it is necessary to previously prepare temperature, pH, concentration of cations and the like. The temperature may be adjusted by using a Peltier element, a heating wire or the like. The temperature may be, for example, about 37 ° C., and the pH may be about 7.

  The cleaning solution may be allowed to flow from the wash flow channel 32 to the hybridization flow channel 37 by opening the open / close switch 36 after the completion of the hybridization. This makes it possible to identify the nucleic acid multiple times using this chip 3 for nucleic acid analysis. When the cleaning fluid flows from the wash flow path 32, the air pump 91 is operated via the air pump control unit 64 and the cleaning fluid is alternately poured. The washing solution flows toward the downstream of the wash flow channel 32 and flows to the hybridization flow channel 37 as it is. The washing solution reaching the hybridization channel 37 flows all remaining nucleic acid and feeds it to the waste 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. 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 portable terminal 5 via the communication unit 62.

  When the portable terminal 5 receives the above data via the communication unit 62, the portable terminal 5 performs nucleic acid identification by analyzing the data using the medical examination application stored in itself, and performs gene analysis. . The form terminal 5 displays the analysis result to the user, or displays various suggestions for future health maintenance to the user. The portable terminal 5 may further transmit the data received through 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 unit 8 and confirmed by a doctor or the like. In addition, findings obtained by analyzing information by the central control unit 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 the collected material from any of the human body, excrement, and soil with the rod-like body 9 and collects it in the collection cartridge 22 in which the liquid L is immersed. The rod-shaped body 9 with the object 95 attached is inserted. In addition, the operation of mounting 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 inlet 33 of the nucleic acid analysis chip 3 In all cases, genetic analysis will be performed automatically.

  For this reason, even a user who does not have specialized knowledge about medical care can perform genetic analysis by himself with extremely simple work.

  In addition, since the pretreatment kit 2 for gene analysis, the chip 3 for nucleic acid analysis, and the chip measurement device 4 do not have complicated apparatus configurations, they can be miniaturized and are suitable for self-examination at home. It can be purchased by individuals because it has an inexpensive structure and can be provided inexpensively.

  Furthermore, according to the present invention, nucleic acids can be extracted with high efficiency from collected products, and in order to identify nucleic acids with high accuracy, serum, bacteria and viruses are removed in advance through the gene analysis pretreatment kit 2 These operations can be carried out extremely easily without any special skill or knowledge.

  In addition, the washing solution can be washed by flowing the washing solution from the wash flow channel 32 to the hybridization flow channel 37, and the nucleic acid analysis chip 3 can be used repeatedly many times, so the cost performance is also very excellent. This cleaning operation can be performed extremely easily.

  Moreover, while aiming at simplification of the device configuration as described above, ease of handling, and improvement of cost performance, the accuracy of the biochemical examination can be maintained at the same level as before, so the same level at medical institutions It is possible to realize gene analysis of at home.

  The present invention is not limited to the embodiment described above. The granular body 223 may be configured by pre-enclosing the liquid L in the epidermis made of an elastic material. In such a case, it is not essential to impregnate the liquid L in the storage container 220. When the rod-shaped body 9 having the collected matter attached to the tip is inserted, the collected matter 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-like body 9 breaks the surface of the granular body 223 to cause the liquid L to flow out. This makes it possible to impregnate the end of the rod-like body 9 with the liquid L that has flowed out.

  In addition, the gene analysis system 1 to which the present invention is applied may be configured in a case accommodation type as shown in FIGS. The constituent elements and members of the case-containing type gene analysis system 1 'are the same as those in the above-described embodiment, and thus the same reference numerals are given and description thereof will be omitted.

  This FIG. 11 shows the appearance of the case-containing type gene analysis system 1 '. In the case-type gene analysis system 1 ', one end is joined by a hinge (not shown), and the case 100a and the case 100b can be opened and closed relative to each other with the hinge (not shown) as a rotation center. A handle 101 is provided on the upper surfaces of the cases 100a and 100b, and a lock mechanism 102 is provided. Moreover, the display 103 and the touch panel 104 may be provided on the outside of the case-containing type 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-described chip measurement device 4 is disposed on the case 100 a side, a personal computer (PC) 50 is connected to the chip measurement device 4, and a battery 105 is further connected to the PC 50. The PC 50 operates with the power supplied from the battery 105. The measurement result by the chip measuring device 4 is sent to the PC 50. The PC 50 is responsible for the same function as the portable terminal 5 described above.

  A nucleic acid analysis chip 3 is disposed on the case 100 b side, and a pretreatment main 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. This enables the measurement described above. An outlet 236 of the pretreatment main body 23 is directly connected to the sample inlet 33 of the chip 3 for nucleic acid analysis, and the liquid L is sent. The pretreatment body 23 and the nucleic acid analysis chip 3 may be arranged in a plurality of sets. A sample can be introduced from the sample introduction port 108 through the branch tube 111 to the pretreatment body 23. The configuration of the sample introduction port 108 is similar to that of the collection cartridge 22. The liquid L from the collection cartridge 22 is sent to the pretreatment body 23 via the branch tube 111. Incidentally, it is assumed that the motive power at the time of sending the liquid L from the collection cartridge 22 to the branch tube 111 is performed via the pump 109. Since the inside of the case 100a, 100b is covered with a reinforced plastic or a reinforced protective film except the sample introduction port 108, scattering of the sample can be prevented.

  According to such a case-containing type gene analysis system 1 ', the system can be compactly integrated in a case such as a suitcase. In addition, high-risk viruses and diseases (HIV, Marburg virus, Ebola virus, smallpox, Lassa fever, etc.) can be measured rapidly without having to outsource them to special management facilities each time. We can quickly detect the epidemic etc. and can control the spread of damage. In particular, according to the case-containing type gene analysis system 1 ′, the rod-shaped body 9 to which the collected matter 95 is attached is only inserted into the sample introduction port 108, and the rest can be measured fully automatically. It can be further improved, and it can be disposed of without any human intervention from the start to the end of the measurement, and therefore it is excellent in terms of hygiene.

  Moreover, according to the present invention, biological substances such as peptides, exosomes, biomarkers, antibodies and the like can be measured in the same way in addition to nucleic acid measurement. In such a case, the liquid L injected through the discharge port 236 in the pretreatment body 23 is allowed to flow directly to the hybridization channel 37 without nucleic acid amplification in the PCR channel 34. The other steps are the same as those of the embodiment described above. That is, when biological substances such as peptides, exosomes, biomarkers, and antibodies are also measured in the same manner, as an alternative to the PCR channel 34, an injection channel for flowing the simply injected liquid L to the hybridization channel 37 It should be done.

  Further, according to the present invention, the first filter 234 for separating serum may be configured, for example, in a form as shown in FIG. Fig. 13 (a) is a side view of the first filter 234, and Fig. 13 (b) is a plan view thereof.

  The first filter 234 includes a membrane filter 301 that constitutes a base, and a separation filter 302 stacked on the membrane filter 301.

  For example, a membrane of HF 135 (capillary flow rate 135 sec / cm, manufactured by Millipore) may be used as the membrane filter 301.

  The separation filter 302 is made of either a hollow fiber or a porous material. Table 7 and Table 8 below show examples of the material constituting the separation filter 302. Tables 7 and 8 below describe further upper concepts of the specific examples. The invention is not limited to the specific example as long as it is included in the broader concept, and any other material may be applied.

  By using the membrane filter 301 made of such a material and the separation filter 302, plasma or serum can be more suitably separated.

  In addition to this, the separation filter 302 may be configured in two or more layers as shown in FIG. Alternatively, the separation filter 302 may be stacked on a local region on the membrane filter 301 in plan view as shown in FIG. In the example of FIG. 13B, the separation filter 302 is stacked in alignment with the right end of the membrane filter 301 in plan view. Since the separation filter 302 is formed only in a local region of the membrane filter 301 in plan view, a boundary line 302 a in plan view with respect to the membrane filter 301 is formed. The end 302 a ′ of the boundary line 302 a in a plan view may have an arc shape.

  Further, FIG. 14 shows another configuration example of the separation filter 302. The shape in plan view of the separation filter 301 stacked in the upper layer of the membrane filter 301 is substantially circular. That is, the shape of the boundary line 302 a in plan view with respect to the membrane filter 301 of the separation filter 302 is semicircular.

  It is assumed that the membrane filter 301 is formed in a rectangular shape in plan view, and its width is 5 mm. Further, it is assumed that the length from the right end in plan view of the separation filter 302 to the boundary 302 a is 10 mm. When 50 ml of blood is dropped from above onto such separation filter 302, the plasma or serum in the dropped blood is directed in the direction of the arrow shown in FIG. Flow toward the boundary line 302a. Then, the plasma or serum that has reached the boundary line 302 a is separated by flowing through the membrane filter 301. It has been confirmed that about 10 to 15 mm of plasma or serum is emitted if the amount of blood dropped is as described above.

  At this time, plasma or serum that has flowed in the separation filter 302 by capillarity collides with the boundary line 302a. When the pressure at the time of the collision is large, the red blood cells are destroyed based on the pressure and so-called hemolysis occurs. In such a case, as shown in FIG. 13 (b), the pressure at the time of the collision is dispersed by forming the end 302a 'of the boundary 302a in an arc shape, so that the pressure itself that is actually loaded is Can be reduced, which in turn can prevent hemolysis. Even in the example shown in FIG. 14, hemolysis can be prevented similarly.

  Further, as shown in FIG. 15, at least the upper end of the boundary line 302 a may be arc-shaped in side view. Also in this case, by dispersing the pressure at the time of the collision described above, it is possible to reduce the pressure itself that is actually loaded, and it is thus possible to prevent hemolysis.

  The separation filter 302 may be added with an anticoagulant or a biological substance. When the sample is blood, the anticoagulant is EDTA (ethylenediaminetetraacetic acid), heparin sodium, citric acid, oxalic acid, sodium fluoride and the like. By adding the anticoagulant, it is possible to prevent the coagulation of the blood itself by mixing it with the anticoagulant in the filter for separation 302 immediately after dropping the blood onto the first filter 234, and hence a good measurement. Is possible.

  Above all, fibrinogen is a substance involved in blood coagulation, which may cause clogging of the test apparatus or interfere with the micro flow path and capillary phenomenon in the microchip constituting the test kit, which makes quantitative measurement difficult, resulting in a minute blood volume. Analysis is difficult.

  For this reason, the following inclusion may be further added to the separation filter 302 in order to remove fibrinogen or to prevent coagulation. Thus, by absorbing or degrading fibrinogen, blood coagulation is prevented and examination is facilitated.

  Examples of the inclusions include hydrophilic monomers such as polyfunctional acrylamide and sulfobetaine monomers and their fine particles, hydroxyapatite (aquamite phosphate), PEG materials (PEG, PLL-g-PEG copolymer, substances to which PEG is added) Etc. may be included. In addition, examples of the inclusion include metal nanoparticles (diameter 5 nm to 500 μm) composed of titanium oxide, gold, silver, zinc oxide, copper, aluminum oxide, titanium oxide etc., fibrinogen decomposition substances (fibrinogen), plasminogen activator (PA) ), Warfarin, acenocoumarol, phenindione, coumarin derivative substances such as anti-vitamin K inhibitors, dabigatran, thrombin inhibitors such as argatroban, rivaroxaban, edoxaban, abixaban, fondaparinux, etc. coagulation factor inhibitors, N-methylol acrylamide 2-hydroxy ethyl methacrylate, hydrophilic polymer fine particles constituted by dispersion polymerization of hydrophilic monomers using acrylic acid as a comonomer, 1,4-cyclohexane dimethanol monoacrylate, N-vinyl pyrrolidone, zeolite, Metal complex (PCP (porous metal complex), copper phthalocyanine, azomethine, etc.), it may be configured with a polyester gel.

  When the sample is DNA (nucleic acid), a DNAase inhibitor may be added as an anticoagulant to prevent DNA degradation by deoxyribonuclease. When the sample is RNA (micro RNA, messenger RNA, etc.), in order to prevent RNA degradation by ribonuclease (endoribonuclease, exoribonuclease, etc.), an RNAse inhibitor is added as an anticoagulant. It is also good. When the sample is saliva, a mucin-degrading enzyme may be added as an anticoagulant in order to degrade mucin, which is a viscosity source in saliva, and to facilitate measurement.

DESCRIPTION OF SYMBOLS 1 gene analysis system 2 pre-processing kit for gene analysis 3 chip for nucleic acid analysis 4 chip measurement device 5 mobile terminal 6 public communication network 7 server 8 central control device 9 rod-like body 12 public communication network 21 plunger 22 cartridge for collection 23 for pretreatment Body 26 Laminate 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 section 42 Detection section 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 Collectable material 96 Protrusion portion 211 Inner cylinder portion 212 pusher 213 rubber part 220 storage container 2 0a Upper chamber 220b Lower chamber 221 Inner wall 222 Projection 223 Granular body 224 Mesh body 225 Film 229 Insertion port 231 Cartridge storage portion 232 Separation portion 234 First filter 235 Second filter 236 Discharge port 237 Cap 301 Membrane filter 302 for separation Filter 302a border 302a 'end

Claims (5)

In a separation filter for separating at least serum or plasma from liquid containing collected material from any of human body, excrement and soil,
Membrane filter,
And a separation filter laminated on the membrane filter,
The separation filter is composed of either a hollow fiber or a porous material, and is composed of two or more layers, and is laminated on a local region on the membrane filter in a plan view, and the plan view for the membrane filter The separation filter according to claim 1 , wherein at least the upper end of the boundary line is arc-shaped in side view . The separation filter is composed of two or more layers, and is laminated on a local region on the membrane filter in a plan view, and the boundary of the membrane filter in a plan view has a circular end at least in a plan view. The separation filter according to claim 1, wherein the separation filter is arc-shaped or substantially semicircular. The separation filter according to claim 1 or 2, wherein an anticoagulant or a biological substance is added to the separation filter. In a pretreatment kit for gene analysis for extracting a biological substance from a collected material collected from any of human body, excrement and soil,
When a plurality of granular bodies are filled, a liquid is immersed in the granular bodies, and when a rod-like body having the collected matter attached to the tip is inserted, the tip is brought into contact with the granular bodies. A collection cartridge for collecting the product and impregnating the liquid with the liquid;
Discharging means for discharging the liquid sent out from the sampling cartridge to the outside;
At least plasma or serum is separated from the liquid delivered from the attached collection cartridge , and a membrane filter and a membrane filter, laminated on either of the hollow fibers and porous material, are separated. A separation filter having an air filter, and a pretreatment body having the discharge means,
The pre-treatment kit for gene analysis, wherein the discharge means discharges the liquid having passed through the separation filter to the outside. The pretreatment body further includes a second filter for separating at least virus and bacteria from the liquid that has passed through the separation filter, and the discharge port allows the liquid that has passed through the second filter to go outside. The preparation kit for gene analysis according to claim 4, wherein the preparation is excreted.

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