Solution supplying device, detection set and detection method

Abstract

A solution supply device (60) supplies a plurality of solutions (first through third solutions (98a)-(98c)) including a reaction liquid to an examination kit (18) having a flow channel that transports the reaction liquid and is provided on a substrate formed from a resin, a solid-phase part provided in the flow channel and on which antibodies (51) are solid-phased, a detection part for detecting reaction of the reaction liquid with the antibodies (51), and a microrelief structure that has a plurality of projections and is formed integrally with the substrate in a region where the flow channel is provided. The solution supply device (60) has a solution unit (80) having a plurality of solution accommodating parts (85), a communicating unit (70) having a plurality of communicating parts (77) provided respectively for the solution accommodating parts (85) and communicating the inside and outside of the solution accommodating parts (85) to supply the solutions to the examination kit (18), and a positioning part for arranging the communicating parts (77) in prescribed positions for communicating the solution accommodating parts (85).

Description

[Technical field]

[0001] The present invention relates to a solution supplying device that supplies a solution to a detection device that detects a target substance in a liquid sample, a detection set, and a detection method. [Background technology]

[0002] In recent years, point-of-care test (POCT) reagents that use antigen-antibody reactions to test for infectious diseases and pregnancy, and measure blood glucose levels, have been attracting attention. Tests and measurements using POCT reagents can determine results in a short time. In addition, POCT reagents are easy to use and inexpensive. Because of these characteristics, POCT reagents are often used for examinations when symptoms are mild and for regular checkups. POCT reagents will also be important diagnostic tools in home medical care, which is expected to increase in the future.

[0003] In tests or diagnoses using a test kit, which is a type of POCT reagent, a liquid sample such as blood is introduced into the test kit to detect a specific target substance contained in the liquid sample. Immunochromatography is often used as a method for detecting a specific target substance from a liquid sample. In immunochromatography, the liquid sample is dropped onto a membrane carrier provided in the test kit, and as the liquid sample moves on the membrane carrier, the target substance in the liquid sample binds to a labeling substance. The target substance further binds specifically and selectively to a substance fixed in the test kit (hereinafter referred to as the detection substance). The resulting changes in color and weight of the test kit are detected. The detection substance may also be called a reagent.

[0004] Nitrocellulose membranes are often used as membrane carriers for moving liquid samples (see Patent Document 1 below). Nitrocellulose membranes have many fine pores with diameters of about several μm, through which liquid samples move by capillary force.

[0005] However, since nitrocellulose membranes are derived from natural products, the pore size and the connection between the pores in the membrane are not uniform, so the flow rate of the liquid sample through the membrane varies depending on the membrane. If there is a difference in the flow rate, the time required to detect the target substance also changes. As a result, it may be erroneously determined that the target substance is not detected before it binds to the label or reagent.

[0006] In order to solve the above problems, a method for artificially creating a microchannel for a liquid sample has been devised (see Patent Documents 2 and 3 below). By using this method, a membrane carrier having a uniform structure can be produced. As a result, it is possible to reduce the possibility of making a mistaken decision that the target substance is not detected before it binds to a labeling substance or a reagent. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] JP 2014-062820 A [Patent Document 2] Patent No. 4597664 [Patent Document 3] Special Publication No. 2012-524894 Summary of the Invention [Problem to be solved by the invention]

[0008] In the immunochromatography method, a liquid sample is dropped onto a membrane carrier provided in a test kit, and as the liquid sample moves on the membrane carrier, the detectable substance in the liquid sample binds to a labeling substance. The detectable substance then specifically and selectively binds to a detection substance immobilized in the test kit. The resulting changes in color and weight of the test kit are then detected. A well-known method for detecting a detectable substance is to detect a color change in the detection zone caused by the detectable substance bound to a labeling substance such as colored latex particles, fluorescent particles, or metal colloid particles binding to a reagent immobilized in the detection zone using an optical measuring device such as an absorbance meter (color change detection method). There is also a method for detecting the concentration of a biomarker by converting it into the concentration of an electrochemically active substance (electrochemical immunochromatography method).

[0009] In the electrochemical immunochromatography method, it is necessary to develop multiple types of solutions such as reaction solution, washing solution, and secondary reaction solution, which increases the labor required for use and increases the detection time, which has been an obstacle to the widespread use of test kits using electrochemical immunochromatography. In other words, there has been a demand for POCT reagents (test kits) that can determine results in a short time, are easy to use, and are inexpensive. Even when a color change detection method is used, the same problem occurs when multiple types of solutions need to be developed.

[0010] The present invention has been made in consideration of the above circumstances, and aims to provide a technology that can eliminate the labor required during use and shorten the detection time when developing multiple types of solutions such as a reaction solution, a washing solution, and a secondary reaction solution in a testing technology using immunochromatography. [Means for solving the problem]

[0011] The present invention provides the following techniques. [1] A solution supplying device that supplies a plurality of solutions including a reaction solution to a detection device having a flow path provided on a substrate formed of resin and transporting the reaction solution, a solid-phase portion in which an antibody or an antigen is solidified and provided in the flow path, a detection portion that detects a reaction of the reaction solution to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in a region in which the flow path is provided, a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; a positioning unit that positions the communication unit at a predetermined position where the communication unit communicates with the solution storage unit; A solution supplying device having the following structure. [2] The solution supplying device described in [1], wherein the communication portion is a tube having one end formed into a needle shape, and when the one end is inserted into the solution storage portion, the solution stored in the solution storage portion passes through the tube and is supplied to the detection device. [3] the communication unit functions as the positioning unit and has a plurality of arrangement parts for accommodating the plurality of solution storage parts, respectively; The communication portion is provided in each arrangement portion, The solution supplying device according to [1] or [2], wherein when the solution storage section is stored in the placement section, the communication section connects the inside and outside of the solution storage section to supply the solution to the detection device. [4] The solution supplying device according to any one of [1] to [3], wherein the communication unit has an adjustment unit for adjusting the position at which the solution is supplied to the detection device. [5] A solution supplying device described in any one of [1] to [4], wherein at least one of the plurality of solution storage sections is capable of being filled with a solution, and the remaining solution storage sections are filled with a predetermined solution in advance. [6] A solution supplying device described in any one of [1] to [5], wherein a single operation of positioning the communication part at a predetermined position to connect the solution storage parts causes a plurality of the solution storage parts in the solution unit to be simultaneously connected by the communication part corresponding to each of them. [7] A solution supplying device that supplies a plurality of solutions including a reaction solution to a detection device having a flow path provided on a substrate formed of resin and transporting the reaction solution, a solid-phase portion in which an antibody or an antigen is solidified and provided in the flow path, a detection portion that detects a reaction of the reaction solution to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in a region in which the flow path is provided, a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; having The solution supply device is configured such that, when an operation is performed once to connect the communication portion to the solution storage portion, all of the solution storage portions are simultaneously connected and the supply of the solution is initiated, wherein the solution unit and the communication unit are configured such that, [8] The communication portion is a tube having one end formed into a needle shape, and when the one end is inserted into the solution storage portion, the solution stored in the solution storage portion passes through the tube and is supplied to the detection device. [7] A solution supplying device as described in. [9] the communication unit has a plurality of arrangement sections each accommodating a plurality of the solution storage sections, The communication portion is provided in each arrangement portion, The solution supplying device according to [7] or [8], wherein when the solution storage section is stored in the placement section, the communication section connects the inside and outside of the solution storage section to supply the solution to the detection device.

[10] The solution supplying device according to any one of [7] to [9], wherein the communication unit has an adjustment unit for adjusting the position at which the solution is supplied to the detection device.

[11] A solution supplying device described in any one of [7] to

[10] , wherein at least one of the plurality of solution storage sections is capable of being filled with a solution, and the remaining solution storage sections are filled with a predetermined solution in advance.

[12] a detection device including: a flow path provided on a substrate formed of resin, for transporting a reaction solution; a solid-phase portion provided in the flow path, in which an antibody or an antigen is solidified; a detection portion for detecting a reaction of the reaction solution to the antibody or the antigen; and a fine concave-convex structure having a plurality of convex portions formed integrally with the substrate in a region in which the flow path is provided; A solution supplying device according to any one of [1] to

[11] , which supplies a plurality of solutions including the reaction solution to the detection device; A detection set having:

[13] The detection set according to

[12] , wherein the detection section is provided on the other end side of the flow path relative to the solid phase section.

[14] The fine uneven structure is a first concave-convex portion in which the plurality of convex portions are relatively sparsely provided; a second concave-convex portion in which the plurality of convex portions are relatively densely arranged; having The detection set according to

[12] or

[13] , wherein the first uneven portion and the second uneven portion are provided closer to one end of the flow path than the solid phase portion.

[15] The detection set according to

[14] , wherein the first uneven portion is provided closer to one end of the flow path than the second uneven portion.

[16] The detection set according to

[14] or

[15] , further comprising a buffer region at the boundary between the first concave-convex portion and the second concave-convex portion where no convex portion is provided.

[17] A step or a slope is provided at a boundary between the first uneven portion and the second uneven portion, The detection set according to any one of

[14] to

[16] , wherein an area of ​​the step or the slope on the side of the first uneven portion is higher than an area on the side of the second uneven portion.

[18] The detection set according to any one of

[14] to

[17] , having a recessed region in which a recess is provided at the boundary between the first recessed portion and the second recessed portion.

[19] The detection set described in any one of

[14] to

[18] , wherein when the first uneven portion and the second uneven portion are adjacent to each other, a ratio (P1 / P2) of a pitch (P1) between the convex portions in the first uneven portion to a pitch (P2) between the convex portions in the second uneven portion is 1.1 or more and 5 or less.

[20] The detection set according to any one of

[12] to

[19] , having a region in which the convex portions are arranged in a diamond lattice pattern. [twenty one] The detection set according to any one of

[12] to

[20] , having a region in which the convex portions are arranged in a regular lattice pattern. [twenty two] The detection set according to any one of

[12] to

[21] , wherein the convex portion is formed as a cone. [twenty three] An introduction part for introducing the solution into the flow channel is further provided, the solution introduced into the flow channel is made up of a plurality of types of solutions, The detection set according to any one of

[12] to

[22] , wherein the introduction section is provided at a plurality of positions corresponding to the plurality of types of solutions. [twenty four] The detection unit is provided with an electrode unit, The detection set according to any one of

[12] to

[23] , wherein the detection unit detects a reaction of the reaction solution with the antibody or the antigen based on a current flowing through the electrode unit. [twenty five] The electrode portion is formed on a convex portion of the fine uneven structure, and the maximum peak height Rp of the roughness curve of the electrode portion is 0.005 μm or more and 10 μm or less, and the average length RSm of the roughness curve element is 0.01 μm or more and 15 μm or less. The detection set described in

[24] .

[26] The detection set according to

[24] or

[25] , wherein the electrode portion has a conductive film layer formed on the convex portions of the fine uneven structure by at least one of sputtering, vacuum deposition, laser ablation, and CVD of a conductive material.

[27] The detection set according to any one of

[24] to

[26] , wherein the electrode portion has a printed layer of a paste containing conductive particles on the convex portions of the fine uneven structure.

[28] The electrode unit has a working electrode and a counter electrode spaced apart from the working electrode, The detection set according to any one of

[24] to

[27] , wherein the working electrode is provided at the same position as the counter electrode or upstream of the counter electrode in the flow path direction.

[29] The detection set according to

[28] , wherein the counter electrode is provided across the entire width of the flow channel.

[30] The detection set according to

[28] or

[29] , wherein the working electrode is provided across the entire width of the flow channel.

[31] The detection set according to any one of

[28] to

[30] , wherein the working electrode is configured as an interdigital electrode.

[32] The detection set according to any one of

[28] to

[31] , wherein the electrode portion further has a reference electrode.

[33] A detection method for detecting a reaction of a liquid sample with an antibody using a detection set described in any one of

[12] to

[32] . Effect of the Invention

[0012] According to the present invention, in a testing technique using immunochromatography, it is possible to provide a technique that can reduce the effort required during use and shorten the detection time when developing multiple types of solutions such as a reaction solution, a washing solution, and a secondary reaction solution. [Brief description of the drawings]

[0013] [Figure 1] FIG. 2 is a top view of the test kit according to the first embodiment. [Diagram 2] FIG. 2 is a top view of the membrane carrier of the first embodiment. [Diagram 3] FIG. 2 is a diagram showing a microstructure and protrusions of the first embodiment. [Figure 4] FIG. 2 is a perspective view of a protrusion according to the first embodiment. [Diagram 5] FIG. 2 is an enlarged view showing boundaries of regions in the microstructure of the first embodiment. [Figure 6] FIG. 2 is a chart showing an example of a testing method using the test kit of the first embodiment. [Figure 7] FIG. 4 is a top view of a membrane carrier according to a second embodiment. [Figure 8] FIG. 11 is an enlarged view showing the boundaries of the regions of the microstructure of the second embodiment. [Figure 9] 13A to 13C are diagrams showing examples of a backflow prevention structure for a solution in a microstructure according to a third embodiment. [Figure 10] FIG. 2 is a photograph of the test piece of Example 1. [Figure 11] FIG. 1 shows images of a test piece in Example 1 taken 30, 140, and 310 seconds after the start of the test. [Figure 12] FIG. 13 is a graph plotting the RGB component ratios at measurement points versus elapsed time in Example 1. [Figure 13] FIG. 13 is a graph showing the results of calculating the mixing ratio of each solution from the results of analyzing the data in FIG. 12 in Example 1. [Figure 14] FIG. 13 is a graph showing the results of calculating the mixing ratio of each solution under the condition that nitrocellulose is used instead of the imprinted sheet of Comparative Example 2. [Figure 15] FIG. 1 is a diagram showing the configuration of a test piece according to a second embodiment. [Figure 16] FIG. 1 is a graph showing the test results of Example 2. [Figure 17] FIG. 13 is a graph showing the fluorescence intensity of Example 2. [Figure 18] 1 is an example of a fluorescent photograph of Example 2. [Figure 19] FIG. 2 is a diagram showing an example of the arrangement of electrode parts in a two-electrode system according to the first embodiment. [Figure 20] 3A and 3B are diagrams illustrating an example of an arrangement of electrode parts in a three-electrode system according to the first embodiment. [Figure 21] FIG. 13 is a diagram showing the configuration of a test piece according to Example 3. [Figure 22] FIG. 13 is a graph showing the test results of Example 3. [Figure 23] FIG. 11 is a chart showing the timing of dropping the solution in Example 4. [Figure 24] FIG. 13 is a graph showing the test results of Example 4. [Diagram 25] FIG. 13 is a cross-sectional view that illustrates a schematic configuration of an inspection set according to a fourth embodiment. [Figure 26] FIG. 13 is a plan view of a solution unit according to a fourth embodiment. [Figure 27] FIG. 13 is a plan view of a communication unit according to a fourth embodiment. [Figure 28] 13A to 13C are diagrams illustrating a solution supplying method using the solution supplying device in the inspection set of the fourth embodiment. [Figure 29] 13A to 13C are diagrams illustrating a solution supplying method using the solution supplying device in the inspection set of the fourth embodiment. [Diagram 30] 13A to 13C are diagrams illustrating a solution supplying method using the solution supplying device in the inspection set of the fourth embodiment. [Diagram 31] FIG. 13 is a cross-sectional view that illustrates a schematic configuration of a solution supplying device according to a first modified example of the fourth embodiment. [Diagram 32] FIG. 13 is a cross-sectional view that illustrates a schematic configuration of a solution supplying device according to a second modified example of the fourth embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] <<First embodiment>> In the following, an embodiment of the present invention will be described. <Overview of the test kit> Fig. 1 is a plan view of a test kit 18 according to this embodiment. Fig. 2 is a plan view showing a schematic diagram of a membrane carrier 3. Fig. 3 shows the fine structure (also called a "fine uneven structure") of the membrane carrier 3 and the convex portions 8 constituting it. Fig. 4 shows a perspective view (SEM image) of the convex portions 8.

[0015] The test kit 18 has a function of detecting a target substance in a liquid sample. Although details will be described later, the test kit 18 is a type of POCT reagent. A liquid sample such as blood is introduced into the test kit 18, and a specific target substance contained in the liquid sample is detected. Immunochromatography is used as a method for detecting a specific target substance from a liquid sample. In this embodiment, a test kit 18 using an electrochemical immunochromatography method for detecting the concentration of a biomarker by converting it into the concentration of an electrochemically active substance will be described.

[0016] As described above, in electrochemical immunochromatography, it is necessary to develop multiple types of solutions (liquid samples) such as a reaction solution, a washing solution, and a secondary reaction solution, which generally increases the labor required for use and lengthens the detection time. In this embodiment, the flow path 2 of the membrane carrier 3 is divided into multiple areas (here, three areas: the first to third microstructure regions 31 to 33), and the flow rate of the solution is controlled to be different for each area. As a structure for making the flow rate different, a microstructure formed on the membrane carrier 3, i.e., a structure that causes a capillary action that determines the speed at which the solution is transported, is set for each area. This will be explained in detail below.

[0017] <Details of Test Kit 18> 1, the test kit 18 includes a membrane carrier 3 and a housing 18a that houses the membrane carrier 3. In this figure, the direction from the upstream on the left side to the downstream on the right side of the figure will be described as the solution traveling direction d (also referred to as the "flow path direction").

[0018] The surface of the membrane carrier 3 has, from the left in the figure, a washing solution zone 3x into which a washing solution is dropped, a liquid droplet zone 3z into which a liquid sample is dropped, and a detection zone 3y for detecting a substance to be detected in the liquid sample. Although not shown here, an absorbent pad for absorbing excess solution is provided downstream of the membrane carrier 3 (to the right in the figure).

[0019] The cleaning solution zone 3x is exposed at the first opening 18b of the housing 18a. The droplet zone 3z is exposed at the third opening 18d of the housing 18a. The detection zone 3y is exposed at the second opening 18c of the housing 18a. The cleaning solution may be dropped at the droplet zone 3z, in which case the first opening 18b may be omitted. When there are multiple types of solutions, inlets (openings) are provided according to the solutions. That is, inlets are provided according to what solution is to be moved, at what timing, and at what speed. Multiple solutions may be dropped at a certain inlet, and the timing may be the same or different.

[0020] An electrode unit 20 is provided in the detection zone 3y for detection by an electrochemical detection method. The electrode unit 20 is, for example, a two-electrode (two-electrode type) consisting of a working electrode 25 on the upstream side of the traveling direction d and a counter electrode 26 on the downstream side. Note that the electrode unit 20 may be a three-electrode type having a reference electrode 27, as described later. A measuring device 21 is connected to the electrode unit 20. The measuring device 21 may be a general measuring device, or may be configured as a device in which a predetermined application is installed on a mobile terminal such as a smartphone.

[0021] <Details of Membrane Carrier 3> As shown in Fig. 2, the membrane carrier 3 is provided with at least one flow path 2 for transporting a liquid sample. As shown in Fig. 3, a microstructure 7 is provided on the bottom surface of the flow path 2. In this embodiment, the microstructure 7 is provided over the entire surface of the membrane carrier 3, and the entire surface of the membrane carrier 3 functions as the flow path 2 for the liquid sample.

[0022] Fig. 3(a) is a top view of the microstructure 7, and Fig. 3(b) is a perspective view of the protrusions 8 that constitute the microstructure. The microstructure 7 is the sum of the protrusions 8. In other words, the membrane carrier 3 has a flat portion 9 that corresponds to the bottom surface of the flow channel 2 for the liquid sample, and a plurality of protrusions 8 protruding from the flat portion 9. By capillary action, the spaces between the multiple protrusions 8 function as flow paths 2 that transport the liquid sample along the surface of the membrane carrier 3. In other words, by capillary action, the voids in the microstructure 7 function as flow paths 2 that transport the liquid sample along the surface of the membrane carrier 3. The multiple protrusions 8 are formed regularly aligned on the surface of the membrane carrier 3, such as in a lattice arrangement (e.g., a diamond lattice arrangement or a regular lattice arrangement), or with translational symmetry.

[0023] The protrusions 8 are, for example, pyramidal, and here, as shown in Fig. 3(b) and Fig. 4, are conical. Alternatively, they may be pyramidal, or may have a shape with the top of the pyramid cut off (truncated pyramid). In any case, it is sufficient that the microstructure 7 formed by the protrusions 8 generates capillary action and transports the liquid sample.

[0024] The microstructure 7 creates a capillary action that transports the liquid sample through the microstructure 7 from the wash zone 3x or the droplet zone 3z on the left side of the figure towards the detection zone 3y (along the travel direction d in FIG. 2).

[0025] 2, in this embodiment, the membrane carrier 3 is divided into three areas from the left: a first microstructure region 31 (first uneven portion), a second microstructure region 32 (second uneven portion), and a third microstructure region 33 (third uneven portion). The first microstructure region 31, the second microstructure region 32, and the third microstructure region 33 have different microstructures 7, and as a result, the speed at which the solution is transported differs for each area.

[0026] The speed at which a solution is transported can be understood from the Poiseuille equation, which explains the flow between parallel plates. For example, in a microstructure 7 that generates capillary action, such as a structure in which multiple protrusions 8 are arranged, the narrower the distance 5 between the protrusions 8, the faster the solution is transported. In other words, the speed for each area can be controlled by appropriately setting the density of the microstructure (the arrangement of the protrusions 8 shown in Figure 3).

[0027] FIG. 5 shows the microstructure 7 as viewed from above. FIG. 5(a) shows the boundary region (first boundary 41) between the first microstructure region 31 and the second microstructure region 32. FIG. 5(b) shows the boundary region (second boundary 42) between the second microstructure region 32 and the third microstructure region 33. In all regions, the protrusions 8 are provided with the same shape and size. In FIG. 5, the protrusions 8 are conical, with a base diameter of 30 μm and a height of 30 μm. As shown in the figure, the arrangement (degree of density) of the protrusions 8 is the sparsest in the first microstructure region 31 on the left side of the figure, and the densest in the third microstructure region 33 on the right side. In other words, the distance 5 between the protrusions 8 in the first microstructure region 31 is the widest, and the distance 5 between the protrusions 8 in the third microstructure region 33 is the narrowest. In the illustrated example, the distance 5 between the convex portions 8 in the first microstructure region 31 is 25 μm, the distance 5 between the convex portions 8 in the second microstructure region 32 is 15 μm, and the distance 5 between the convex portions 8 in the third microstructure region 33 is 2 μm.

[0028] When the substance to be detected in the liquid sample reaches the detection zone 3y, it is detected as a current value by the measuring device 21 through the electrode unit 20 (working electrode 25, counter electrode 26) provided in the detection zone 3y. That is, a potential difference is applied between the working electrode 25 and the counter electrode 26 of the electrode unit 20, and the oxidation current is measured by the measuring device 21. When the color change detection method is used, the substance to be detected is detected by the color change of the detection zone 3y.

[0029] <Materials for Membrane Carrier 3> The membrane carrier 3 including the microstructure 7 (multiple convex portions 8) is made of, for example, a thermoplastic. That is, the membrane carrier 3 having the microstructure 7 can be produced by processing a film-shaped substrate made of a thermoplastic by thermal imprinting. The thermoplastic constituting the membrane carrier 3 may be, for example, at least one selected from the group consisting of polyester-based resins, polyolefin-based resins, polystyrene-based resins, polycarbonate-based resins, fluorine-based resins, and acrylic-based resins. A specific thermoplastic may be, for example, at least one of polyethylene terephthalate (PET), cycloolefin polymer (COP), polypropylene (PP), polystyrene (PS), polycarbonate (PC), polyvinylidene fluoride (PVDF), and polymethyl methacrylate (PMMA).

[0030] The glass transition point Tg or melting point Tm of the thermoplastic may be 80 to 180° C. The storage modulus of the thermoplastic at a temperature 20° C. higher than the glass transition point Tg is 1.0 Pa or more and 1.0×10 7 The storage modulus of a thermoplastic plastic at a temperature 20° C. higher than the melting point Tm may be 1.0 Pa or more and 1.0×10 7 Pa or less. The glass transition or melting of a thermoplastic occurs at a temperature below 80°C, and the storage modulus of the thermoplastic at a temperature 20°C higher than the glass transition point or melting point is 1.0 x 10 7If the resistivity is less than 1 Pa, it is practically difficult to use a thermoplastic plastic as a solid at room temperature, and it is difficult to prepare a membrane carrier by thermal imprinting. If the glass transition or melting of the thermoplastic occurs at a temperature higher than 180° C., the molding temperature during thermal imprinting becomes high, and the productivity of the membrane carrier decreases. In other words, if the temperature required to soften the thermoplastic during thermal imprinting is higher than 180° C., the productivity of the membrane carrier decreases. The storage modulus of a thermoplastic at a temperature 20°C higher than the glass transition point or melting point is 1.0 x 10 7 When the pressure is equal to or less than 1 Pa, the molding pressure required for producing the microstructure can be kept small, and the microstructure can be produced under relatively mild conditions, improving production efficiency.

[0031] The cone-shaped (here, circular cone) protrusions 8 can be formed by thermal imprinting using a mold. When forming the cone using a mold, the volume of metal to be cut out from the surface of the metal member during mold fabrication is significantly reduced compared to when a mold is used to form a groove-shaped flow path (line and space structure), and the processing cost of the mold is reduced. In contrast, when fabricating a mold for forming a line and space structure, a large amount of metal must be cut out from the metal member.

[0032] In addition, the top of the pyramid is thinner than the base of the pyramid, so when forming the pyramid using a mold, the volume of metal that is cut out from the surface of the metal member during mold creation is significantly reduced compared to when a cylinder having the same base as the pyramid is formed using a mold, and the processing costs of the mold are reduced.

[0033] Furthermore, the porosity of a microstructure in which the pyramids are regularly aligned is greater than that of a line-and-space structure. Also, the porosity of a microstructure in which the pyramids are regularly aligned is greater than that of a structure in which multiple pillars having the same base as the pyramids are regularly aligned. Therefore, the microstructure in which the pyramids are regularly aligned can increase the flow rate of a liquid sample, which is advantageous for detecting a target substance.

[0034] <Shape and dimensions of membrane carrier 3> As described above, the shape of the bottom surface 10 of the cone (protrusion 8) can be freely selected, and may be a cone as shown in Figure 3(b) and Figure 4, or a pyramid (such as a square pyramid or a hexagonal pyramid). For ease of processing the mold and to reduce processing costs, it is desirable for the bottom surface 10 of the cone (protrusion 8) to be circular or polygonal (for example, a square, diamond, rectangle, triangle, or hexagon).

[0035] The diameter 4 of the bottom surface 10 of the protrusion 8 is, for example, 10 to 1000 μm. If the diameter 4 of the bottom surface 10 of the protrusion 8 is smaller than 10 μm, the cost of microfabrication of the mold becomes high, and it is difficult to uniformly fabricate numerous microstructures 7 on the surface of the membrane carrier 3 having a large area. Therefore, a microstructure 7 that is too small is not suitable for practical use. If the diameter 4 of the bottom surface 10 of the microstructure 7 is smaller than 10 μm, the capillary force required to move the liquid sample tends to be weak. If the diameter 4 of the bottom surface 10 of the microstructure 7 is larger than 1000 μm, the volume of the metal cut out from the metal member during the fabrication of the mold becomes large, and the fabrication cost of the mold and the membrane carrier 3 becomes high. If the diameter 4 of the bottom surface 10 of the microstructure 7 is larger than 1000 μm, the area of ​​the flow path 2 in the membrane carrier 3 must also be increased, and the test kit 18 becomes large, which is disadvantageous for the transportation of the test kit 18 itself. When the protrusion 8 (microstructure 7) is a cone, the diameter 4 of the base 10 of the protrusion 8 may be the diameter 4 of the base 10 (circle) of the cone.

[0036] The height 6 of the protrusions 8 is, for example, 10 to 500 μm. If the height 6 of the protrusions 8 is less than 10 μm, the capillary force required to move the liquid sample tends to be weak. If the height 6 of the protrusions 8 is greater than 500 μm, it is difficult to completely fill the thermoplastic plastic into the recesses (recesses corresponding to the shape of the protrusions 8 of the microstructure 7) of the mold during thermal imprinting.

[0037] The overall shape of the membrane carrier 3 is not particularly limited, and may be, for example, a polygon such as a rectangle, a circle, or an ellipse. When the membrane carrier 3 is a rectangle, the vertical width L1 of the membrane carrier 3 may be, for example, 2 to 100 mm, and the horizontal width L2 of the membrane carrier 3 may be, for example, 3 to 100 mm. Furthermore, the horizontal widths L21 to L23 of the first to third microstructure regions 31 to 33 may each be, for example, 1 to 50 mm. The thickness of the membrane carrier 3 excluding the height 6 of the microstructure 7 (i.e., the protrusions 8) may be, for example, 0.1 to 10 mm.

[0038] The aspect ratio Lv / Lh of the protrusion 8 may be 1 / 10 or more and 2 / 1 or less. If the aspect ratio Lv / Lh is smaller than 1 / 10, the contact area between the liquid sample and the flow channel 2 is small, and the capillary force is reduced, so that the liquid sample tends to be difficult to move. If the aspect ratio Lv / Lh is larger than 2 / 1, the productivity of the membrane carrier 3 by thermal imprinting decreases. If the protrusion 8 is a pyramid (more specifically, a cone) as in this embodiment, the length Lh of the protrusion 8 in the horizontal direction may be the diameter 4 of the bottom surface 10 of the protrusion 8. In addition, the length Lv of the protrusion 8 in the vertical direction may be the height 6 of the protrusion 8 from the flat portion 9 of the membrane carrier 3.

[0039] The ratio D2 / D1 of the diameter 4 (D1) of the bottom surface of the convex portion 8 to the closest center distance (D2) between the convex portions 8 may be greater than 1 and less than or equal to 5. The ratio D2 / D1 cannot be less than 1. If the ratio D2 / D1 is greater than 5, the contact area between the liquid sample and the flow channel 2 decreases, the capillary force decreases, and the liquid sample tends to be difficult to move. When the convex portion 8 is a cone as in this embodiment, the diameter 4 (D1) of the bottom surface 10 of the convex portion 8 may be the diameter of the bottom surface of the cone, and the closest center distance D2 may be the distance between the apexes of a pair of adjacent convex portions 8 (cones). The diameter 4 (D1) of the bottom surface 10 of the convex portion 8 may be the same as the length Lh of the convex portion 8 in the horizontal direction described above. Therefore, the aspect ratio Lv / Lh may be expressed as Lv / D1.

[0040] In addition, when comparing the pitch (distance between apexes) between the convex portions 8 of the fine concave-convex structure between adjacent zones, the ratio (P1 / P2) between the pitch P1 of the zone with the fine concave-convex structure that is relatively sparse and the pitch P2 of the zone with the fine concave-convex structure that is relatively dense is 1.1 or more and 5 or less. In this case, if the pitch between the convex portions 8 of the first fine structure region 31 is P11, the pitch between the convex portions 8 of the second fine structure region 32 is P21, and the pitch between the convex portions 8 of the third fine structure region 33 is P23, the ratio (P11 / P21) is 1.1 or more and 5 or less, and the ratio (P21 / P23) is 1.1 or more and 5 or less. The ratio (P1 / P2) is set depending on the speed at which the solution is desired to move. Regarding the lower limit of the ratio (P1 / P2), if it is small, there will be no speed difference between the zones, and the significance of providing a difference in the degree of coarseness and density of the fine concave-convex structure will be reduced. From this viewpoint, the ratio (P1 / P2) is preferably 1.2 or more, more preferably 1.3 or more. If the upper limit is too large, the difference in the solution movement speed between the zones becomes too large, making it difficult to adjust the speed throughout the test kit 18. From this viewpoint, the ratio is preferably 4 or less, more preferably 3 or less.

[0041] <Electrode placement> An example of the arrangement of the electrode section 20 will be described with reference to FIGS. 19(a) to 19(c) show examples of the arrangement of the working electrode 25 and the counter electrode 26 when the electrode unit 20 is of a two-electrode type. The working electrode 25 and the counter electrode 26 are provided apart from each other. Here, the working electrode 25 is provided at the same position as the counter electrode 26 or upstream of the counter electrode 26 with respect to the traveling direction d. The working electrode 25 may be configured as, for example, a comb-shaped electrode.

[0042] In the arrangement example shown in FIG. 19(a), the working electrode 25 is provided over the entire width of the flow path 2. The counter electrode 26 is provided over the entire width of the flow path 2 in a region downstream of the working electrode 25 by a predetermined distance. The working electrode 25 and the counter electrode 26 are rectangular in top view, but are not limited to this shape and may be various shapes such as an ellipse or a semicircle. The working electrode 25 and the counter electrode 26 are provided so as to block the width of the flow path 2, but are not limited to this and may be provided only in a partial region in the width direction as shown in FIG. 1 and FIG. 2. The width of both or either one of the working electrode 25 and the counter electrode 26 may be shortened.

[0043] 19(b), the counter electrode 26 is provided in a U-shape with a concave upstream side when viewed from above. Furthermore, the working electrode 25 is provided in a rectangular shape in the concave region of the counter electrode 26. The most upstream position of the working electrode 25 and the upstream position of the counter electrode 26 are in the same position.

[0044] In the arrangement example shown in FIG. 19(c), a working electrode 25 and a counter electrode 26, each of which has a rectangular shape, are provided symmetrically in the width direction.

[0045] 20(a) to 20(f) show examples of the arrangement of the working electrode 25, the counter electrode 26, and the reference electrode 27 when the electrode unit 20 is of a three-electrode type.

[0046] 20(a), the working electrode 25, the reference electrode 27, and the counter electrode 26 are arranged side by side from the upstream side to the downstream side, and are provided across the entire width direction of the flow channel 2. This can also be said to be a configuration in which the reference electrode 27 is arranged between the working electrode 25 and the counter electrode 26 in the arrangement shown in FIG.

[0047] In the arrangement example shown in FIG. 20(b), the width of the reference electrode 27 in the arrangement shown in FIG. 20(a) is shortened and provided in a rectangular shape in the center of the width direction of the flow channel 2.

[0048] 20(c), a working electrode 25 and a counter electrode 26 are provided from the upstream side on the left side of the flow path 2 as shown in the figure, and a reference electrode 27 is provided on the right side of the flow path 2. The reference electrode 27 is provided in an elongated shape in the traveling direction d from a position on the upstream side of the working electrode 25 to a position on the downstream side of the counter electrode 26.

[0049] In the arrangement example shown in FIG. 20(d), the working electrode 25 and the counter electrode 26 are arranged in the same manner as in FIG. 19(b), and further, a reference electrode 27 is provided upstream of the working electrode 25 on the left side in the figure.

[0050] In the arrangement example shown in FIG. 20(e), the concave right end of the counter electrode 26 in the arrangement shown in FIG. 19(b) is shortened toward the downstream side, and a reference electrode 27 is provided in the shortened region.

[0051] 20(f), the counter electrode 26 is provided across the entire width of the flow channel 2. Furthermore, a working electrode 25 is provided on the left side of the flow channel 2, and a reference electrode 27 is provided on the right side, so as to be symmetrical. <Electrode configuration> The electrode section 20 (working electrode 25 and counter electrode 26 in the case of a two-electrode system, and further a reference electrode 27 in the case of a three-electrode system) may be formed by providing a conductive material on the protrusions 8 of the microstructure 7. The conductive material is not particularly limited, but examples thereof include gold, silver, platinum, palladium, carbon, graphene, carbon nanotubes (CNTs), and composites thereof. The reference electrode 27 is not particularly limited, but examples thereof include an Ag / AgCl electrode.

[0052] The conductive material of the protrusions 8 may be, for example, a conductive film formed by at least one of sputtering, vacuum deposition, laser ablation, and CVD (chemical vapor deposition), or a printed layer formed by inkjet printing, screen printing, or the like using a paste (ink) containing conductive particles. The working electrode 25 may be surface-modified with thiol or the like for antibody immobilization.

[0053] In this case, the maximum peak height Rp of the roughness curve of the electrode part 20 is 0.005 μm or more and 10 μm or less, and the average length RSm of the roughness curve element is 0.01 μm or more and 15 μm or less. By making the surface roughness in this manner, a good capillary force can be generated, and the surface area of ​​the electrode part 20 is increased, so that the amount of the signal obtained can be increased. These surface roughnesses can be calculated by analyzing SEM images such as those shown in FIG. 4.

[0054] <Manufacturing method of test kit 18> The manufacturing method of the test kit 18 is obtained by the following steps. Process 1 (Thermal imprint process) The method includes a step of applying the surface of a metal mold, on which multiple recesses are formed, to a film-shaped substrate made of a thermoplastic plastic and heating the substrate to produce a film carrier 3 having a microstructure (multiple protrusions 8) corresponding to the shape of the recesses. The method for producing the test kit 18 further includes a step of fixing a reagent or labeling substance to the detection zone 3y on the surface of the membrane carrier 3 having the microstructure 7, more specifically, to the solid phase portion 50 of the third microstructure region 33. The microfabrication method of the mold used in the thermal imprinting process may be, for example, etching, photolithography, mechanical cutting, laser processing, etc. A microfabrication method suitable for the processing size and processing range can be selected.

[0055] It is desirable to perform a mold release treatment before performing thermal imprinting. In the mold release treatment, for example, a monolayer may be formed on the mold surface to reduce the surface energy. As a result, the membrane carrier 3 made of a thermoplastic plastic can be easily peeled off from the mold surface after thermal imprinting.

[0056] The thermal imprinting method may be either a flat plate press method or a roll method. In the flat plate press method, a mold is placed on a base material made of a thermoplastic plastic between upper and lower stages facing each other in parallel, and these are sandwiched between the stages. Then, the mold and base material are heated and pressurized through the stage. Such a flat plate press method is excellent in terms of molding accuracy. The roll method is a method in which a heated roll type mold is used to mold by the sandwiching pressure between rolls. The roll method is excellent in productivity.

[0057] The conditions for thermal imprinting, such as molding temperature, molding pressure, and transfer time, may be selected according to the size of the microfabrication, the shape of the microstructure (protrusions 8), the size of the processing area, and the like. For example, in the case of a flat plate press type, the molding temperature may be a temperature 20 to 50°C higher than the glass transition point Tg, or a temperature 20 to 50°C higher than the melting point Tm. The molding pressure may be 1 to 10 MPa. The transfer time (the time during which the mold and substrate are held under pressure) may be 3 to 10 minutes. Thermal imprinting under the above conditions facilitates accurate transfer of the microstructure of the mold to the surface of the substrate.

[0058] Depending on the type of thermoplastic constituting the membrane carrier 3 and the type of reagent (detection substance), it may be difficult to fix the reagent (detection substance) to the solid phase part 50 of the membrane carrier 3. In this case, by performing an appropriate surface treatment in advance only on the detection zone 3y, it becomes easier to fix the reagent (detection substance) to the detection zone 3y of the membrane carrier 3 (i.e., the solid phase part 50).

[0059] The surface treatment method for the detection zone 3y is not limited in any way, and may be various methods such as various plasma treatments, UV treatments, UV / ozone treatments, or surface modification with 3-Aminopropyltriethoxysilane or Glutaraldehyde.

[0060] The reagent (detection substance) immobilized in the detection zone 3y may be, for example, an antibody. For example, in Fig. 2, an antibody is immobilized on the solid phase part 50 of the third microstructure region 33. The solid phase part 50 is provided upstream of the electrode part 20 in the traveling direction d of the solution.

[0061] The antibody is a substance that causes an antigen-antibody reaction with the substance to be detected. The antibody may be a polyclonal antibody or a monoclonal antibody. The substance to be detected is not limited in any way, and may be any substance capable of causing an antigen-antibody reaction with the antibody, such as various pathogens and various clinical markers. Specific examples of the substance to be detected may be virus antigens such as influenza virus, norovirus, adenovirus, respiratory syncytial virus, HAV, HBs, and HIV. The substance to be detected may be bacterial antigens such as MRSA, group A streptococcus, group B streptococcus, and Legionella, and toxins produced by bacteria. The substance to be detected may be hormones such as mycoplasma, chlamydia trachomatis, and human chorionic gonadotropin. The substance to be detected may be C-reactive protein, myoglobin, cardiac troponin, various tumor markers, agricultural chemicals, and environmental hormones. In particular, the test kit 18 according to this embodiment is particularly useful when it is necessary to detect a substance to be detected, such as influenza virus, norovirus, C-reactive protein, myoglobin, and cardiac troponin, and to take urgent measures to treat diseases caused by these substances. The substance to be detected may be an antigen capable of inducing an immune reaction by itself. The substance to be detected may be a hapten that cannot induce an immune reaction by itself but can bind to an antibody through an antigen-antibody reaction with the antibody.

[0062] <Testing method using Test Kit 18> The testing method using the test kit 18 will be described with reference to the chart shown in Fig. 6 and the above-mentioned Figs. 1 to 5. In Fig. 6, attention is focused on the third microstructure region 33 of the microstructure 30.

[0063] S1: Device preparation process First, the test kit 18 and the solutions to be used (reaction solution, washing solution, secondary reaction solution) are prepared. As described above, the antibody 51 is fixed to the solid phase portion 50 of the third microstructure region 33.

[0064] S2: Reaction solution development process When the reaction liquid is dropped from the liquid droplet zone 3z onto the second microstructure region 32, it moves to the third microstructure region 33 due to the capillary action of the microstructures 7. The detection target 91 and the detection target (labeled substance) 92 in the reaction solution are immobilized by reacting with the antibody 51. Excess reaction solution is absorbed by the water-absorbing pad, but a part of the detection target 91a and the detection target (labeled substance) 92a is not immobilized and remains on the third microstructure region 33.

[0065] S3: Cleaning solution development process Next, a cleaning solution 93 is dropped from the cleaning solution zone 3x to wash the detection target 91a and the detection target (labeled substance) 92a that are not fixed to the solid phase portion 50 and remain in the third microstructure region 33. The detection target (labeled substance) 92a has an alkaline phosphatase (ALP) label.

[0066] S4: Secondary reaction solution development process After washing, the secondary reaction liquid (e.g., p-aminophenyl phosphate 94) dropped onto the second microstructure region 32 moves to the third microstructure region 33 by the capillary action of the microstructure 7. The p-aminophenyl phosphate 94 reacts with the analyte (label) 92 immobilized on the antibody 51 to generate an electrically active substance (here, p-aminophenol 95). This substance is correlated (proportional) to the amount (concentration) of the analyte (label) 92 immobilized on the antibody 51. Therefore, the concentration of the analyte can be measured accurately and stably from the value of the oxidation current measured by the electrode section 20.

[0067] As described above, according to this embodiment, in the test kit 18 to which the immunochromatography method is applied, the speed at which the solution moves in the flow channel 2 of the microstructure 7 of the membrane carrier 3 (the speed due to the capillary action) can be set to a plurality of regions with different speeds. As a result, even when it is necessary to develop a plurality of types of solutions such as a reaction solution, a cleaning solution, and a secondary reaction solution, the timing at which these solutions are developed can be adjusted according to the mode of use. Therefore, the time and effort required for timing adjustment, etc., which would otherwise be required, can be eliminated, and a stable and appropriate test can be performed. Specifically, by using a predetermined tool (solution supply device), etc., each solution can be dropped simultaneously at a plurality of different locations taking into consideration the timing at which each solution is developed. In other words, each solution can be developed with only one operation. The configuration using the tool (solution supply device) will be described in the fourth embodiment described later.

[0068] <<Second embodiment>> The test kit of this embodiment will be described with reference to Figures 7 and 8. The difference from the first embodiment is in the structure of the membrane carrier 103, and mainly the different points will be described, and the description of the same configurations and functions will be omitted as appropriate.

[0069] Fig. 7 is a plan view showing a schematic view of the membrane carrier 103. Fig. 8 shows enlarged images of the boundaries between adjacent regions. Fig. 8(a) is an image of a first boundary 141 between the first microstructure region 131 and the second microstructure region 132. Fig. 8(b) is an image of a second boundary 142 between the second microstructure region 132 and the third microstructure region 133. Fig. 8(c) is an image of a third boundary 143 between the third microstructure region 133 and the fourth microstructure region 134.

[0070] As shown in the figure, the membrane carrier 103 has a rectangular shape with a predetermined vertical width L10 and horizontal width L20. From the left, the membrane carrier 103 comprises a first microstructure region 131 (horizontal width L201), a second microstructure region 132 (horizontal width L202), a third microstructure region 133 (horizontal width L203), and a fourth microstructure region 134 (horizontal width L204). As in the first embodiment, these regions have different densities of protrusions in the microstructure, and as a result, the speed due to capillary action is different.

[0071] Specifically, the first microstructure region 131 is the coarsest (region A11), the third microstructure region 133 is the second coarsest (region A13), the second microstructure region 132 is the third coarsest (region A12), and the fourth microstructure region 134 is the densest (region A14). Also, a solid phase portion 150 is provided in the fourth microstructure region 134.

[0072] In addition, a buffer region of a predetermined width L31 is provided at the second boundary 142 between the second fine structure region 132 and the third fine structure region 133. No fine structure (i.e., a convex portion) is provided in the buffer region. Similarly, a buffer region of a predetermined width L32 is provided at the third boundary 143 between the third fine structure region 133 and the fourth fine structure region 134. By providing such a buffer region, it is possible to absorb the difference in the transport amount of the solution for each region and prevent the occurrence of backflow, etc. For example, between the second fine structure region 132 and the third fine structure region 133, the microstructure of the third fine structure region 133 on the downstream side is coarse. Therefore, the movement speed of the solution is higher in the second fine structure region 132. As a result, if there is no buffer region at the second boundary 142, backflow may occur depending on the amount of solution developed. However, by providing a buffer region in which no capillary force is generated as in this embodiment, it is possible to prevent the occurrence of backflow due to the movement speed of the solution or the amount developed.

[0073] <<Third embodiment>> In this embodiment, six examples of the backflow prevention structure for a solution will be described with reference to Fig. 9. Note that, although the cross-sectional views of some regions of the configurations corresponding to the above-mentioned membrane carriers 3 and 103 are extracted and described here, the same can be applied to other regions.

[0074] In the membrane carrier 203 shown in FIG. 9(a), a step 241 is provided at the boundary between a first fine structure region 231 and a second fine structure region 232 such that the second fine structure region 232 side is lower.

[0075] 9(b), a sloping portion 341 that becomes lower in the downstream direction is provided at the boundary between a first microstructure region 331 and a second microstructure region 332. The sloping portion 341 may be a buffer region without a protrusion, or may be a fine uneven structure having a protrusion.

[0076] 9(c), an inclined portion 441 that becomes lower in the upstream direction is provided at the boundary between the first microstructure region 431 and the second microstructure region 432. The boundary between the inclined portion 441 and the second microstructure region 432 is a step portion 442.

[0077] In the membrane carrier 503 shown in FIG. 9( d ), a recess 541 is provided at the boundary between the first microstructure region 531 and the second microstructure region 532 .

[0078] In the membrane carrier 603 shown in FIG. 9(e), the first microstructure region 631 and the third microstructure region 633 are formed horizontally, but the second microstructure region 632 has an inclination that becomes lower toward the downstream side.

[0079] 9(f), the first microstructure region 731, the second microstructure region 732, and the third microstructure region 733 all have an inclination that decreases toward the downstream side. Here, a configuration in which all of the regions have the same inclination angle is shown, but the inclination angle may be different for each region.

[0080] The configurations of Figs. 9(a) to (f) can be appropriately combined to form a desired membrane carrier, and an optimal flow path and solution movement speed can be realized according to the type and amount of solution to be developed.

[0081] <<Fourth embodiment>> The test set 1 of this embodiment will be described with reference to Figures 25 to 32. In this embodiment, a solution supplying device 60 is used as a solution supplying tool to carry out a test method using a test kit 18 (membrane carrier 3 having antibody 51 immobilized thereon), which is the test device described in the first to third embodiments, to simplify the effort of supplying a solution in carrying out the test method. Since the configuration of the test kit 18 (membrane carrier 3) is similar to that described in the first to third embodiments, the solution supplying device 60 will be mainly described below.

[0082] <Test set 1> Fig. 25 is a cross-sectional view showing a schematic configuration of the test set 1. Fig. 26 is a plan view of the solution unit 80. Fig. 27 is a plan view of the communication unit .

[0083] The test set 1 includes the above-mentioned test kit 18 and a solution supplying device 60 that supplies each solution to the test kit 18. The solution supplying device 60 contains a plurality of solutions (first to third solutions 98a to 98c in this case) used in the test. As the solutions, the solutions shown in the test method of the first embodiment (see FIG. 6) can be used. Specifically, the first solution 98a can be a reaction solution, the second solution 98b can be a cleaning solution, and the third solution 98c can be a secondary reaction solution.

[0084] <Solution supply device 60> The solution supplying device 60 comprises a solution unit 80 and a communication unit 70, and is arranged to cover the upper surface of the test kit 18. By operation of a user (here, a person in charge of testing), the above solutions (first to third solutions 98a to 98c) are supplied to specified areas of the membrane carrier 3 of the test kit 18. The details will be explained below.

[0085] <Solution Unit 80> The solution unit 80 has a plate 82 that is rectangular in plan view, and a plurality of solution containers 85 that are recessed in the plate 82 .

[0086] The plate 82 and the solution storage section 85 are integrally made of a resin material. For example, thermoplastics such as polypropylene and polyethylene, which are exemplified as the material of the membrane carrier 3, can be used as the resin material. The thickness of the plate 82 and the solution storage section 85 is set so that the solution unit 80 has a certain rigidity, but the thickness of the bottom surface 86 is set so that the communication unit 70 having a syringe needle structure can pierce it.

[0087] As shown in Figs. 25 and 26, the solution storage section 85 has first to third solution storage sections 85a to 85c in this order from the left side of the figure. Here, the first solution storage section 85a stores a first solution 98a. The second solution storage section 85b stores a second solution 98b. The third solution storage section 85c stores a third solution 98c. When the communication unit 70 and the solution unit 80 are stacked, the first to third solution storage sections 85a to 85c are set so as to fit into first to third arrangement sections 75a to 75c of the communication unit 70 described below.

[0088] The solution storage section 85 (first to third solution storage sections 85a to 85c) has a cylindrical shape with a bottom. The size of the solution storage section 85 is appropriately set according to the amount of solution to be stored. The shape of the solution storage section 85 is not limited to a cylindrical shape, and may be a square tube shape. The inner diameter of the solution storage section 85 is set to a size that can store the arrangement section 75 and is approximately the same as the outer diameter of the arrangement section 75. The depth of the solution storage section 85 is set so that a bottom surface 86 of the solution storage section 85 abuts against a bottom surface 76 of the arrangement section 75 when the solution storage section 85 (first to third solution storage sections 85a to 85c) is fitted into the arrangement section 75 (first to third arrangement sections 75a to 75c) described later.

[0089] <Communication unit 70> The communication unit 70 includes a plate 72 having a rectangular shape in a top view, a frame 71 extending downward from the periphery of the plate 72, a plurality of arrangement portions 75 recessed in the plate 72, and a communication portion 77 provided in the arrangement portion 75. The frame 71, the plate 72, and the arrangement portion 75 are integrally formed from a resin material. As with the solution unit 80, for example, a thermoplastic plastic such as polypropylene or polyethylene can be used as the resin material. The thicknesses of the frame 71, the plate 72, and the arrangement portion 75 are set so that the communication unit 70 has a certain rigidity.

[0090] The frame 71 functions to separate the plate 72 and the test kit 18 by a predetermined distance, and also functions to appropriately fix the communication unit 70 to the test kit 18. As a fixing function, a configuration can be adopted in which a lower end portion 73 of the frame 71 fits into a step portion 19 provided on the periphery of the test kit 18.

[0091] 25 and 27, the arrangement portion 75 includes first to third arrangement portions 75a to 75c, which are arranged in this order from the left side of the figure. When the communication unit 70 and the solution unit 80 are stacked, the first to third arrangement portions 75a to 75c are set so as to fit into the first to third solution storage portions 85a to 85c of the solution unit 80.

[0092] The arrangement section 75 (first to third arrangement sections 75a to 75c) has a cylindrical shape with a bottom. The size of the cylindrical shape is set so that the solution accommodation section 85 (first to third solution accommodation sections 85a to 85c) can be fitted and accommodated therein. The shape of the arrangement section 75 is not limited to a cylindrical shape and may be a square tube shape as long as the solution accommodation section 85 can be fitted and accommodated therein. The inner diameter of the arrangement section 75 is set to be approximately the same as the outer diameter of the solution accommodation section 85. The depth of the arrangement section 75 is set so that the bottom surface 86 of the solution accommodation section 85 abuts against the bottom surface 76 of the arrangement section 75 when the arrangement section 75 is fitted into the solution accommodation section 85 (first to third solution accommodation sections 85a to 85c).

[0093] The communication parts 77 (first to third communication parts 77a to 77c) have a structure of a tube (straight tube) with an end (needle tip 79) on the upper side in the figure formed in a needle shape, that is, an injection needle. Here, the bottom surface 76 of the arrangement part 75 is vertically communicated so that the needle tip 79 of the injection needle is on the inside side of the arrangement part 75. When the arrangement part 75 is fitted into the solution storage part 85, the communication part 77 pierces the bottom surface 86 of the solution storage part 85. In other words, the arrangement part 75 (first to third arrangement parts 75a to 75c) and the solution storage part 85 (first to third solution storage parts 85a to 85c) function as a positioning part that positions the communication parts 77 (first to third communication parts 77a to 77c) at a predetermined position that communicates with the solution storage part 85 (first to third solution storage parts 85a to 85c). As a result, the solution contained in solution container 85 passes through communication section 77 and drips onto test kit 18 .

[0094] The communicating part 77 is made of a resin material or a metal, similar to a general injection needle. Thermoplastics such as polypropylene and polyethylene can be used as the resin material. Stainless steel can be used as the metal. The size of the communicating part 77 is not particularly limited as long as it can perforate the bottom surface 86 of the solution storage part 85 and smoothly drip the solution, but the inner diameter of the communicating part 77 can be, for example, 1 to 2 mm.

[0095] The position on the test kit 18, more specifically, the position on the membrane carrier 3, to which the solution is dropped is determined by the position of the solution storage section 85, i.e., the position of the communication section 77. Therefore, the inlets (first opening 18b to third opening 18d) described in the first embodiment (see FIG. 1, etc.) may be provided as a single opening rather than being provided separately. Furthermore, the solution supply device 60 (particularly the communication unit 70) may function as the housing 18a of the test kit 18. Furthermore, the number of communication portions 77 for each arrangement portion 75 is not limited to one, but may be multiple, and may differ for each arrangement portion 75.

[0096] <Solution supply method (testing method)> 28 to 30, a solution supplying method using the solution supplying device 60 in the inspection set 1 will be described. The first to third solutions 98a to 98c start to be supplied simultaneously by a single operation by the user.

[0097] 28, the communication unit 70 is attached to the test kit 18. At this time, the frame lower end portion 73 of the communication unit 70 fits into the step portion 19 of the test kit 18, thereby positioning and attaching the communication unit 70 to the test kit 18. The first to third solution storage portions 85a to 85c of the solution unit 80 store first to third solutions 98a to 98c.

[0098] 29, the solution unit 80 is placed on the communication unit 70, and the first to third solution storage sections 85a to 85c are fitted into the first to third arrangement sections 75a to 75c. At this time, the first to third communication sections 77a to 77c are in contact with the bottom surfaces 86 of the first to third solution storage sections 85a to 85c but are not perforated.

[0099] 30, the user operates to push the solution unit 80 into the communication unit 70. With this single operation, the first to third communication parts 77a to 77c simultaneously perforate the bottom surfaces 86 of the first to third solution storage parts 85a to 85c, thereby communicating the inside and outside of the first to third solution storage parts 85a to 85c. As a result, the first to third solutions 98a to 98c stored in the first to third solution storage parts 85a to 85c start to be simultaneously supplied to the membrane carrier 3 of the test kit 18.

[0100] As described above, according to this embodiment, it is possible to start supplying a plurality of solutions (first to third solutions 98a to 98c) to the test kit 18 simultaneously with a single operation. That is, when developing a plurality of types of solutions such as a reaction solution, a cleaning solution, and a secondary reaction solution, it is possible to save the user's trouble during use and shorten the detection time. In addition, it is possible to keep the timing of developing a plurality of types of solutions constant.

[0101] <Modification 1 of the solution supply device 60> Not limited to the above configuration, various configurations can be adopted for the solution supplying device 60. Modifications of the solution supplying device 60 will be described with reference to Figs.

[0102] The solution supplying device 60 of the first modification shown in FIG. 31 has a different shape of the communication part 77 of the communication unit 70. Specifically, the first communication part 77a has a structure (adjustment part) for adjusting the position where the first solution 98a is supplied to the test kit 18. As a structure of the adjustment part, the first communication part 77a not only extends vertically but also extends horizontally. In the figure, the first communication part 77a is bent once toward the second communication part 77b on the right side of the figure and extends, and finally bent vertically toward the test kit 18 side. This adjusts the drip position (development position) of the first solution 98a. In addition, by adopting a structure that allows the first communication part 77a to pivot around the mounting position of the placement part 75 as an axis, further position adjustment is possible. Although an example has been shown in which the communication portion 77 having the adjustment portion structure is applied to the first communication portion 77a, it may also be applied to the second communication portion 77b or the third communication portion 77c, and may be applied to any of the communication portions 77.

[0103] <Modification 2 of solution supply device 60> The solution supplying device 60 of the second modification shown in FIG. 32 differs from the solution supplying device 60 of the first modification shown in FIG. 31 in that the solution unit 80 has a different configuration. At least one of the multiple solution containing sections 85 of the solution unit 80 can be filled with a solution, and the remaining solution containing sections 85 are filled with a predetermined solution in advance. Specifically, the first and second solution containing sections 85a and 85b are filled with a first solution 98a and a second solution 98b in advance, respectively, and the upper openings are covered with lids 87a and 87b. On the other hand, the upper opening of the third solution containing section 85c is not covered, and can be filled with a third solution 98c during inspection.

[0104] Although the embodiments (first to fourth embodiments) of the present invention have been described above with reference to the drawings, these are merely examples of the present invention, and various configurations (modifications) other than those described above can also be adopted. For example, the flow path 2 is provided as a fine structure (fine uneven structure) of the membrane carrier 3 on a substrate formed of resin, but if a fine structure (fine uneven structure) with different densities can be provided depending on the region of the flow path 2, various configurations, materials, etc. can be adopted.

[0105] <Summary of the embodiment> The features of the present invention can be briefly summarized as follows. (1) The solution supplying device of the present embodiment is a solution supplying device that supplies a plurality of solutions including the reaction liquid to a detection device having a flow path provided on a substrate formed of resin and transporting a reaction liquid, a solid-phase portion in which an antibody or an antigen is solidified and provided in the flow path, a detection portion that detects a reaction of the reaction liquid to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in a region in which the flow path is provided, a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; a positioning unit that positions the communication unit at a predetermined position where the communication unit communicates with the solution storage unit; has. (2) The communication part is a tube with one end formed into a needle shape, and when the one end is inserted into the solution storage part, the solution stored in the solution storage part is supplied to the detection device through the tube. (3) The communication unit functions as the positioning unit and has a plurality of arrangement parts that accommodate the plurality of solution storage parts, respectively; The communication portion is provided in each arrangement portion, When the solution storage section is stored in the placement section, the communication section communicates between the inside and outside of the solution storage section to supply the solution to the detection device. (4) The communication unit has an adjustment unit that adjusts the position at which the solution is supplied to the detection device. (5) At least one of the plurality of solution storage sections can be filled with a solution, and the remaining solution storage sections are filled with a predetermined solution in advance. (6) By performing the operation of positioning the communication parts at predetermined positions for connecting the solution storage parts once, the multiple solution storage parts provided in the solution unit are simultaneously connected by the communication parts corresponding to each of them. (7) The solution supplying device of the present embodiment is a solution supplying device that supplies a plurality of solutions including the reaction liquid to a detection device having a flow path provided on a substrate formed of resin and transporting a reaction liquid, a solid-phase portion in which an antibody or an antigen is solidified and provided in the flow path, a detection portion that detects a reaction of the reaction liquid to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in a region in which the flow path is provided, a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; having The solution unit and the communication unit are configured such that, by performing an operation in which the communication portion communicates with the solution storage portion once, all of the solution storage portions are simultaneously connected and the supply of the solution is initiated. (8) The communication part is a tube having one end formed into a needle shape, and when the one end is inserted into the solution storage part, the solution stored in the solution storage part is supplied to the detection device through the tube. (9) The communication unit has a plurality of arrangement sections each accommodating a plurality of the solution storage sections, The communication portion is provided in each arrangement portion, 9. The solution supplying device according to claim 7, wherein when the solution storage section is stored in the placement section, the communication section provides communication between the inside and outside of the solution storage section to supply the solution to the detection device. (10) The communication unit has an adjustment unit that adjusts the position at which the solution is supplied to the detection device. (11) At least one of the plurality of solution storage sections can be filled with a solution, and the remaining solution storage sections are filled with a predetermined solution in advance. (12) A detection set of the present invention includes a detection device having a flow path provided on a substrate formed of a resin and transporting a reaction solution, a solid-phase portion provided in the flow path in which an antibody or an antigen is solidified, a detection portion for detecting a reaction of the reaction solution to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in the region in which the flow path is provided; The detection device further includes the above-mentioned solution supply device that supplies a plurality of solutions including the reaction liquid to the detection device. (13) The detection section is provided on the other end side (i.e., downstream side) of the flow channel relative to the solid phase section. (14) The fine uneven structure is a first concave-convex portion in which the plurality of convex portions are relatively sparsely provided; a second concave-convex portion in which the plurality of convex portions are relatively densely arranged; having The first concave-convex portion and the second concave-convex portion are provided on one end side (that is, upstream side) of the flow channel relative to the solid phase portion. (15) The first concave-convex portion is provided closer to one end of the flow path (that is, on the upstream side) than the second concave-convex portion. (16) The boundary between the first concave-convex portion and the second concave-convex portion has a buffer region where no convex portion is provided. (17) A step or a slope is provided at a boundary between the first uneven portion and the second uneven portion, The area of ​​the step or the slope on the side of the first uneven portion is higher than the area on the side of the second uneven portion. (18) A recessed region having a recess is provided at the boundary between the first concave-convex portion and the second concave-convex portion. (19) When the first uneven portion and the second uneven portion are adjacent to each other, the ratio (P1 / P2) of the pitch (P1) between the convex portions in the first uneven portion to the pitch (P2) between the convex portions in the second uneven portion is 1.1 or more and 5 or less. (20) The projections have a region in which they are arranged in a rhombic lattice pattern. (21) The projections have a region in which they are arranged in a regular lattice pattern. (22) The convex portion is provided as a cone. (23) An introduction part for introducing the solution into the flow channel, the solution introduced into the flow channel is made up of a plurality of types of solutions, The introduction portion is provided at a plurality of locations corresponding to the plurality of types of solutions. (24) The detection unit is provided with an electrode unit, The detection unit detects the reaction of the reaction solution with the antibody or the antigen based on the current flowing through the electrode unit. (25) The electrode portion is formed on a convex portion of the fine uneven structure, and the maximum peak height Rp of the roughness curve of the electrode portion is 0.005 μm or more and 10 μm or less, and the average length RSm of the roughness curve element is 0.01 μm or more and 15 μm or less. (26) The electrode portion has a conductive film layer on the convex portions of the fine concave-convex structure, the conductive material being formed by at least one of sputtering, vacuum deposition, laser ablation, and CVD. (27) The electrode portion has a printed layer of a paste containing conductive particles on the convex portions of the fine concave-convex structure. (28) The electrode portion has a working electrode and a counter electrode spaced apart from the working electrode, The working electrode is provided at the same position as the counter electrode or upstream of the counter electrode in the flow path direction. (29) The counter electrode is provided across the entire width of the flow channel. (30) The working electrode is provided across the entire width of the flow channel. (31) The working electrode is configured as an interdigital electrode. (32) The electrode portion further includes a reference electrode. (33) The detection method of this embodiment detects the reaction of a liquid sample with an antibody using the above-mentioned detection set. EXAMPLES

[0106] The present invention will be specifically described below with reference to examples (Examples 1 and 2), but the present invention is not limited to these examples. In the following examples, an evaluation experiment was carried out on the flow rate control of a solution when the membrane carrier had a microstructure having a plurality of regions with different densities.

[0107] [Example 1] In this example, an experiment for quantitatively evaluating the state in which three colored aqueous solutions are replaced when they are spread on a membrane carrier will be described.

[0108] 1. Experiment (1) In order to confirm the technology of controlling solution spreading by changing the microstructure, a flow channel 2 was fabricated in polycarbonate (Teijin PC-2151) for the membrane carrier 3 having the configuration shown in Fig. 2 of the first embodiment, in which three regions were connected together with the distances between the protrusions 8 of the first microstructure region 31, the second microstructure region 32, and the third microstructure region 33 being 25 μm, 15 μm, and 2 μm, respectively. The fabrication conditions (thermal imprinting process) were as follows.

[0109] <Thermal imprint process (transfer of fine structures)> The microstructure on the mold surface was transferred to the surface of a film-like substrate made of thermoplastic plastic by the following thermal imprinting process. In the thermal imprinting process, X-300 manufactured by SCIVAX was used. In the thermal imprinting process, the surface of the above-mentioned mold on which the microstructure (multiple recesses) was formed was applied to a film-like substrate made of thermoplastic plastic, and the mold and substrate were heated and pressurized. The molding temperature was 180°C. The applied pressure was 5.5 MPa. The transfer time was 5 minutes. After the transfer of the microstructure, the mold and substrate were cooled to 140°C while pressure was applied to them. After cooling, the pressure was removed. The above thermal imprinting process obtained the membrane carrier of Example 1. This membrane carrier had a surface including multiple cones (microstructures) and flat parts. The shape and size of the convex parts (cones) on the surface of the membrane carrier were consistent with the shape and size of the concave parts (inverted cones) formed on the mold.

[0110] The protrusion 8 has a conical structure with a diameter 4 and a height 6 both of 30 μm. The membrane carrier 3 has a vertical width L1 of 5 mm, and the first to third microstructure regions 31 to 33 have horizontal widths L21 to L23 of 20 mm, respectively.

[0111] (2) A water-absorbing pad used in Navi-Flu was attached to the end of the third microstructure region 33, overlapping it by 5 mm. Furthermore, a conjugate pad used in Navi-Flu was fixed to the point where the distance between the microstructures changes (the position corresponding to the first boundary 41 and the second boundary 42 in Figure 2) to prepare a test specimen. Figure 10 shows a photograph of the prepared test specimen.

[0112] (3) The conjugate pads were numbered "1," "2," and "3" in order of proximity to the absorbent pad, and an aqueous solution with the composition shown in Table 1 was dripped onto each pad. The amount of liquid dripped was determined taking into consideration the distance to the absorbent pad and the amount trapped by the conjugate pad during development. To simplify the operation and result verification of this experiment, the pads were dripped in the order of "1," "2," and "3" every 10 seconds.

[0113] [Table 1]

[0114] (4) The process of each colored solution spreading was recorded on video, and the color change at the midpoint between drop point 1 and the absorbent pad was analyzed by image analysis.

[0115] 2.Results The captured video was imaged every 10 seconds and imported into image analysis software (software name: "Image J"). Images taken 30, 140, and 310 seconds after the start of the test are shown in Figure 11. Figure 11(a) is the image after 30 seconds, Figure 11(b) is the image after 140 seconds, and Figure 11(c) is the image after 310 seconds.

[0116] The measurement point was set at the midpoint between drop point 1 and the absorbent pad, and the RGB display data at that point was recorded. Next, the component ratios of each RGB color were calculated according to formula 1. As an example, the conversion data for each aqueous solution of red (R), green (G), and blue (B) are shown in Table 2. (R or G or B component ratio) = (R or G or B value) / (R value + G value + B value) …Formula 1

[0117] [Table 2]

[0118] A graph plotting the RGB component ratios at the measurement points against the elapsed time is shown in Figure 12. From the results shown in Figure 12, it can be confirmed that the change from green to red to blue over time can be quantified.

[0119] Furthermore, in order to quantitatively evaluate the inspection results and the state of the flow, the following formula 2 was considered as a method for quantitatively evaluating the mixture ratio. This is to determine how much of the single color component ratios in Table 2 should be mixed to achieve the RGB component ratio at each measurement point. The solver function of the spreadsheet software Excel was used to minimize the error ε between the actual measurement value and the calculated value. ε=|R_r x+G_r y+B_r zr|+|R_g x+G_g y+B_g zg|+|R_b x+G_b y+B_b zb| …Formula 2 Find x, y, and z that minimize ε expressed by equation 2.

[0120] Here, R_r, R_g, and R_b respectively indicate the ratios of R, G, and B components in the red solution. G and B have the same meanings for the green and blue solutions. r, g, and b respectively indicate the actual ratios of R, G, and B components at the measurement point. x, y, and z respectively indicate the mixture ratios of the red, green, and blue solutions at the measurement point, and were limited to x+y+z=1. By analyzing the data in FIG. 12 using Equation 2, the mixing ratio of each solution could be expressed as shown in FIG. 13.

[0121] From the above results, the time-dependent change in the mixing ratio of the solutions was quantitatively evaluated by image analysis. In other words, it was confirmed that the speed of the solution movement in the membrane carrier flow channel can be adjusted by appropriately setting the density of the microstructure of each region of the membrane carrier (the spacing of the protrusions), and multiple solutions can be spread.

[0122] The results of a similar test and evaluation carried out using nitrocellulose instead of the imprinted sheet (comparative example) are shown in Figure 14. In the case of the imprinted sheet, the structure (protrusions) is conical, so the solution in the flow channel can be observed from directly above, but the nitrocellulose is in the form of a nonwoven fabric, so only the color change on the outermost surface could be observed. Keep this in mind when considering the results.

[0123] Comparing with FIG. 13 (Example of imprinted sheet), it can be seen that the test time is significantly different. This shows the difference in the development flow rate between the imprinted sheet and nitrocellulose, and it was confirmed that the imprinted sheet with a high flow rate is advantageous in terms of quickly replacing the development solution. The test time can be shortened by adjusting the overall length of the nitrocellulose, but in that case, the risk of the solutions dropping at the same time mixing increases. In fact, in a test in which the overall length and the amount of dropped liquid were halved, the solutions mixed. It is possible to create a device that can be judged in a short time and does not mix the solutions by devising the structure, but the design freedom is low because the flow rate cannot be adjusted. In that respect, when the membrane carrier is composed of an imprinted sheet as shown in Example 1, the freedom of flow rate adjustment is high.

[0124] 3. Summary According to Example 1, in the case of the membrane carrier of the imprinted sheet, the flow rate can be controlled by adjusting the microstructure and material, and it is possible to flexibly respond to market needs.

[0125] [Example 2] 1. Experiment (1) In order to confirm the technology of controlling solution development by changing the microstructure, a flow channel having four regions was fabricated in polycarbonate (Teijin PC-2151) for the membrane carrier 103 having the configuration shown in Fig. 7 of the second embodiment, in which the distances between the apexes of the protrusions 8 in the first microstructure region 131, the second microstructure region 132, the third microstructure region 133, and the fourth microstructure region 134 were 105 μm, 60 μm, 80 μm, and 30 μm, respectively. The fabrication conditions were the same as in Example 1. Fig. 15(a) shows a schematic diagram of the flow channel as seen from the side, and Fig. 15(b) shows a top view (photograph) of the test piece that was actually fabricated. The protrusion 8 has a conical structure with a diameter 4 and a height 6 both of 32 μm. The vertical width L1 of the membrane carrier 103 is 5 mm, the horizontal width L201 of the first microstructure region 131 is 15 mm, the horizontal width L202 of the second microstructure region 132 is 30 mm, the horizontal width L203 of the third microstructure region 133 is 45 mm, and the horizontal width L204 of the fourth microstructure region 134 is 40 mm. In side view, the second microstructure region 132, the third microstructure region 133 and the fourth microstructure region 134 are inclined at an inclination angle of 2.2°. The first boundary 141 is a boundary where the first fine structure region 131 and the second fine structure region 132 are continuous, with no buffer region present. The second boundary 142 has a buffer area (unprocessed area) with a width L31 of 0.15 mm. The third boundary 143 is a buffer area (unprocessed area) having a width L32 of 0.20 mm.

[0126] (2) Experiment 1 (RGB image analysis): In Experiment 1, the color change at a given point was analyzed from RGB components using the same experimental and analytical methods as in Example 1, and the state in which the liquid was replaced was quantitatively confirmed. Specifically, the color change at a point 5 mm from the most downstream point (the right end of the fourth microstructure region 134 in the figure) was analyzed from the RGB components. That is, the mixing ratio of the solutions during development was evaluated based on the ratio of the RGB components of each solution. A graph of the evaluation results is shown in FIG. 16. This corresponds to FIG. 13 in Example 1, and it was confirmed that the components with the largest ratios changed over time from green (G) to red (R) to blue (B).

[0127] (3) Experiment 2 (CRP detection performance evaluation): 15(a), the solution development process was as follows: 10 μL of cleaning solution was dropped onto the fourth microstructure region 134, 10 seconds later 10 μL of CRP solution was dropped onto the third microstructure region 133, 1 minute later 15 μL of fluorescent labeling solution was dropped onto the second microstructure region 132, and finally 2 minutes later 30 μL of cleaning solution was dropped onto the first microstructure region 131. Fluorescence intensity was measured 10 minutes after the last solution (cleaning solution) was dropped.

[0128] The solutions used were as follows: Washing solution: PBS containing 2wt% Triton X-100 CRP solution: A mixture of lavage fluid and CRP solution at a specified concentration Fluorescent labeling solution: A mixture of the washing solution and fluorescently labeled anti-CRP antibody solution to give an antibody concentration of 30 μg / mL.

[0129] FIG. 17 shows the phosphor intensity for each CRP concentration. FIG. 18 shows images of the fluorescence intensity. FIG. 18(a) shows the result when the CRP concentration was 0 ng / mL, and FIG. 18(b) shows the result when the CRP concentration was 10 ng / mL. From the results shown in FIG. 17 and FIG. 18, a CRP concentration-dependent fluorescence intensity was obtained at a CRP concentration of 1 ng / mL or more. From this, it can be seen that by applying the membrane carrier having multiple microstructures as described above to electrochemical detection (electrochemical immunochromatography), it is possible to electrically measure the desired detection target substance.

[0130] [Example 3] 1. Experiment (1) Test piece (membrane carrier 103) In Example 3, in order to make the test system closer to the actual use conditions, the test conditions in Experiment 2 of Example 2 were partially changed, and a CRP detection test was carried out in which human serum was mixed into the sample solution. The structure of the test piece (membrane carrier 103) corresponding to Fig. 7 was prepared in polycarbonate (Teijin PC-2151) with a flow channel having four regions, in which the apex-to-apex distances of the protrusions 8 in the first microstructure region 131, the second microstructure region 132, the third microstructure region 133, and the fourth microstructure region 134 were 100µm, 60µm, 95µm, and 30µm, respectively. The preparation conditions were the same as those in Examples 1 and 2. A schematic diagram of the test piece (flow path) seen from the side is shown in Figure 21. Thirty-six samples were prepared as test pieces. The protrusion 8 has a cone structure with a diameter 4 and a height 6 of 32 μm. The vertical width L1 of the membrane carrier 103 is 5 mm, the horizontal width L201 of the first microstructure region 131 is 36.95 mm, the horizontal width L202 of the second microstructure region 132 is 5 mm, the horizontal width L203 of the third microstructure region 133 is 40 mm, and the horizontal width L204 of the fourth microstructure region 134 is 40 mm. In side view, the second microstructure region 132, the third microstructure region 133 and the fourth microstructure region 134 are inclined at an inclination angle of 2.1°. The first boundary 141 is a boundary where the first fine structure region 131 and the second fine structure region 132 are continuous, with no buffer region present. The second boundary 142 has a buffer area (unprocessed area) with a width L31 of 0.15 mm. The third boundary 143 is a buffer area (unprocessed area) having a width L32 of 0.15 mm.

[0131] (2) Antibody solid phase 1 μL of the anti-CRP antibody solid phase solution was dropped at a position 17.5 mm from the most downstream end of the test piece, and the test piece was dried in an atmosphere at 45° C. for 1 hour to solidify 25 ng of anti-CRP antibody.

[0132] As shown in Figure 21, the flow channel was inclined at 2.1°, and the CRP solution, fluorescently labeled anti-CRP antibody solution (fluorescently labeled anti-CRP antibody concentration in the developing solution: 45 μg / mL), and developing solution (PBS containing 2 wt% Triton X-100) were spread in this order from different dropping points. The composition of the CRP solution used is shown in Table 3. The amount of developing solution and the interval between dropping the solution are shown in Table 4. Each test was performed with n=3. Ten minutes after all the solution had been spread, the absorbent pad and the imprint sheet (membrane carrier 103) were separated to prevent backflow of the solution, and the fluorescence intensity of the antibody solid phase portion was then measured.

[0133] 2.Results The results of measuring the fluorescence intensity of each test are shown in Figure 22. The fluorescence intensity shown here is the value obtained by subtracting the background fluorescence intensity around the antibody solid phase from the fluorescence intensity of the antibody solid phase. Note that the exposure time for measuring the fluorescence intensity was 1 second in Figure 22(a), and 1 / 6 second in Figure 22(b). The same minimum detection sensitivity and measurement range (detection range) of more than three orders of magnitude were achieved regardless of the presence or absence of serum. The fluorescence intensity tended to decrease in the presence of serum, which is presumably because the proteins in serum (up to 80 mg / mL) inhibited the reaction between the antibody and CRP.

[0134] In Example 3, for simplicity, a flow path was designed that functions when one operator drips three types of solutions successively, but the design of this example can be easily modified to a flow path design that functions when three types of solutions are dripped simultaneously using a jig or the like.

[0135] Specifically, to adjust the 6-second time difference between when the CRP solution is dropped and when the fluorescently labeled anti-CRP antibody solution is dropped, the flow path length of the third microstructure region 133 should be lengthened by 6×V mm, assuming that the flow rate in the third microstructure region 133 is V (mm / s). The time difference between when the CRP solution is dropped and when the developing solution is dropped can also be adjusted by lengthening the flow path length of the first microstructure 131 using a similar concept.

[0136] As described above, it is clear that by making minor modifications to the flow path design of Example 3, equivalent detection performance can be obtained even when three types of solutions are dropped simultaneously using a jig or the like.

[0137] In addition, in Examples 2 and 3, a preferable example is shown having a plurality of types of microstructure regions with different flow rates. On the other hand, it is possible to obtain the same effect by applying the solution backflow prevention structure of the third embodiment (see FIG. 9) to a membrane carrier having a single microstructure region. In other words, when the solution backflow prevention structure of the third embodiment is applied to the membrane carrier, it is possible to sequentially develop a plurality of types of solutions in the solid phase portion and the detection portion without controlling the flow rate, and it is possible to obtain detection performance equivalent to Examples 2 and 3. In this case, in order to make the time until the contact of the solutions and the accompanying mixing occur equivalent to Examples 2 and 3, it is necessary to make the flow path length from the most upstream part of the flow path to the first boundary 141, the flow path length from the first boundary 141 to the second boundary 142, and the flow path length from the second boundary 142 to the third boundary 143 longer than those of Examples 2 and 3.

[0138] [Table 3] [Table 4]

[0139] [Example 4] In Example 4, an electrochemical detection test was carried out on an imprinted sheet based on the results of Examples 1 to 3. In this example, from the viewpoint of checking whether electrochemical detection could be performed appropriately, all solutions were dropped at different timings at the same position on the fourth microstructure region 134. 1. Experiment (1) Test piece (membrane carrier 103) A membrane carrier 103 was prepared having the same structure as the test piece produced in Example 3. The vertical width L1 of the membrane carrier 103 was 5 mm, the horizontal width L201 of the first microstructure region 131 was 36.95 mm, the horizontal width L202 of the second microstructure region 132 was 5 mm, the horizontal width L203 of the third microstructure region 133 was 40 mm, and the horizontal width L204 of the fourth microstructure region 134 was 40 mm.

[0140] (2) Electrode part (working electrode, counter electrode) An imprint sheet was attached onto a substrate made of a SUS plate with polyimide tape attached, and gold was vacuum-deposited onto the substrate through a mask processed into an electrode shape to form electrode portion 20. The electrode shapes are as follows: Working electrode: 1mm x 5mm (flow path width) Counter electrode: 3mm x 5mm (channel width) Gap between electrodes: 0.5mm Electrode position: The downstream end of the counter electrode is 15.5 mm from the most downstream end of the flow path.

[0141] (3) Antibody solid phase As in Example 3, 1 μL of the anti-CRP antibody solid phase solution was dropped at a position 5 mm upstream from the working electrode, and dried in an atmosphere at 45° C. for 1 hour to immobilize 25 ng of anti-CRP antibody.

[0142] (4) Measuring equipment The electrodes 20 (working electrode, counter electrode) and the substrate were electrically connected with silver paste (Dotite D-550), and the substrate was clamped with an alligator clip of an electrochemical measurement device (Solartron 1252A) for measurement. A potential of +50 mV was applied between the working electrode and the counter electrode to develop the solution, and the current value was plotted against time (see FIG. 24).

[0143] (5) Solution drip timing and drip solution FIG. 23 is a chart showing the timing of dropping the solution in Example 4. First, 10 μL of the cleaning solution was dropped (first step S11). Two minutes after the first step S11, 10 μL of the CRP and ALP-labeled CRP mixed solution was dropped (second step S12). Furthermore, two minutes after the second step S12, 10 μL of the cleaning solution (4%) was dropped (third step S13). Finally, three minutes after the third step S13, 10 μL of the p-aminophenyl phosphate Na solution was dropped (fourth step S14).

[0144] The solutions dropped were as follows: Washing solution: PBS containing 2wt% Triton X-100 ALP-labeled CRP: Commercially available CRP was labeled with a labeling kit (LK13, Dojindo Laboratories). CRP, ALP-labeled CRP mixed solution: CRP and ALP-labeled CRP are suspended in the washing solution to a specified concentration. Washing solution (4%)…PBS containing 4wt% Triton X-100 p-Aminophenyl phosphate sodium solution: Dissolve p-aminophenyl phosphate sodium in the washing solution to a concentration of 5 mM.

[0145] 2.Results The measurement results are shown in Figure 24. Figure 24(a) shows the results when the CRP concentration of the CRP and ALP-labeled CRP mixed solution was 0 μg / mL and the ALP-CRP concentration was 1.25 μg / mL, and Figure 24(b) shows the results when the CRP concentration of the mixed solution was 12.5 μg / mL and the ALP-CRP concentration was 1.25 μg / mL. As shown in the figure, a current value reflecting the timing of dropping the solution and the CRP concentration was detected.

[0146] This application claims priority based on Japanese Patent Application No. 2021-139004, filed on August 27, 2021, the disclosure of which is incorporated herein in its entirety. [Explanation of symbols]

[0147] 1 Test Set 2 Flow Path 3, 103, 203, 303, 403, 503, 603, 703 Membrane carriers 3x cleaning fluid zones 3y Detection Zone 3z Droplet Zone 8 Convex 18 Test Kits 18a Case 18b First opening 18c Second opening 18d Third opening 20 Electrode part 21 Measuring Equipment 25 Working electrode 26 Opposite 27 Reference pole 31, 131, 231, 331, 431, 531, 631, 731 1st fine structure region 32, 132, 232, 332, 432, 532, 632, 732 second fine structure region 33, 133 Third microstructure region 134 4th fine structure region 41, 141, 241 first boundary 42, 142 second boundary 143 Third boundary 50, 150 solid phase part 51 Antibodies 60 Solution supply device 70 Connecting unit 71 Frames 72 Plate 73 Bottom end of frame 75 Placement section 75a to 75c First to third arrangement sections 76, 86 Bottom 77 Communication part 77a to 77c First to third communication parts 79 Needle tip 80 Solution Units 82 Plate 85 Solution storage section 85a to 85c First to third solution storage units 87a to 87b First and second lid parts 98a-98c Solutions 1-3 341, 441 Slope 541 Recess

Claims

1. A solution supplying device that supplies a plurality of solutions including a reaction solution to a detection device having a flow path provided on a substrate formed of resin and transporting the reaction solution, a solid-phase portion in which an antibody or an antigen is solidified and provided in the flow path, a detection portion that detects a reaction of the reaction solution to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in a region in which the flow path is provided, a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; a positioning unit that positions the communication unit at a predetermined position where the communication unit communicates with the solution storage unit; having The communication portion is a tube with one end formed into a needle shape, and when the one end is inserted into the solution storage portion, the solution stored in the solution storage portion is supplied to the detection device through the tube.

2. A solution supplying device that supplies a plurality of solutions including a reaction liquid to a detection device having a flow path provided on a substrate formed of resin for transporting the reaction liquid, a solid phase portion provided in the flow path in which an antibody or an antigen is solidified, a detection portion that detects a reaction of the reaction liquid to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in the region in which the flow path is provided, a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; a positioning unit that positions the communication unit at a predetermined position where the communication unit communicates with the solution storage unit; having the communication unit functions as the positioning unit and has a plurality of arrangement parts for accommodating the plurality of solution storage parts, respectively; The communication portion is provided in each arrangement portion, When the solution storage section is stored in the placement section, the communication section communicates between the inside and outside of the solution storage section to supply the solution to the detection device.

3. A solution supplying device that supplies a plurality of solutions including a reaction liquid to a detection device having a flow path provided on a substrate formed of resin for transporting the reaction liquid, a solid phase portion provided in the flow path in which an antibody or an antigen is solidified, a detection portion that detects a reaction of the reaction liquid to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in the region in which the flow path is provided, a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; a positioning unit that positions the communication unit at a predetermined position where the communication unit communicates with the solution storage unit; having The communication unit has an adjustment unit that adjusts a position at which the solution is supplied to the detection device.

4. 4. The solution supplying device according to claim 1, wherein at least one of the plurality of solution containing sections is capable of being filled with a solution, and the remaining solution containing sections are filled with a predetermined solution in advance.

5. A solution supplying device as described in any one of claims 1 to 3, wherein the operation of positioning the communication portion at a predetermined position for connecting the solution storage portions is performed once, so that the multiple solution storage portions provided in the solution unit are simultaneously connected by the communication portion corresponding to each of them.

6. A solution supplying device that supplies a plurality of solutions including a reaction solution to a detection device having a flow path provided on a substrate formed of resin and transporting the reaction solution, a solid-phase portion in which an antibody or an antigen is solidified and provided in the flow path, a detection portion that detects a reaction of the reaction solution to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in a region in which the flow path is provided, a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; having the solution unit and the communication unit are configured such that, by performing an operation of connecting the communication parts to the solution storage parts once, all the solution storage parts are simultaneously connected to each other and the supply of the solution is started, The communication portion is a tube with one end formed into a needle shape, and when the one end is inserted into the solution storage portion, the solution stored in the solution storage portion is supplied to the detection device through the tube.

7. A solution supplying device that supplies a plurality of solutions including a reaction liquid to a detection device having a flow path provided on a substrate formed of resin for transporting the reaction liquid, a solid phase portion provided in the flow path in which an antibody or an antigen is solidified, a detection portion that detects a reaction of the reaction liquid to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions integrally formed on the substrate in the region in which the flow path is provided, a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; having the solution unit and the communication unit are configured such that, by performing an operation of connecting the communication parts to the solution storage parts once, all the solution storage parts are simultaneously connected to each other and the supply of the solution is started, The communication unit has an adjustment unit that adjusts a position at which the solution is supplied to the detection device.

8. the communication unit has a plurality of arrangement sections each accommodating a plurality of the solution storage sections, The communication portion is provided in each arrangement portion, The solution supplying device according to claim 6 or 7, wherein when the solution storage section is stored in the placement section, the communication section provides communication between the inside and outside of the solution storage section to supply the solution to the detection device.

9. 8. The solution supplying device according to claim 6, wherein at least one of the plurality of solution containing sections is capable of being filled with a solution, and the remaining solution containing sections are filled with a predetermined solution in advance.

10. a detection device including: a flow path provided on a substrate formed of resin, for transporting a reaction solution; a solid-phase portion provided in the flow path, in which an antibody or an antigen is solidified; a detection portion for detecting a reaction of the reaction solution to the antibody or the antigen; and a fine concave-convex structure having a plurality of convex portions formed integrally with the substrate in a region in which the flow path is provided; A solution supplying device according to claim 1 , which supplies a plurality of solutions including the reaction solution to the detection device; A detection set having:

11. The detection set according to claim 10 , wherein the detection section is provided on the other end side of the flow channel relative to the solid phase section.

12. The fine uneven structure is a first concave-convex portion in which the plurality of convex portions are relatively sparsely provided; a second concave-convex portion in which the plurality of convex portions are provided relatively densely; having The first uneven portion and the second uneven portion are provided on one end side of the flow channel relative to the solid phase portion. The detection set according to claim 10.

13. The detection set according to claim 12 , wherein the first uneven portion is provided closer to one end side of the flow channel than the second uneven portion.

14. The detection set according to claim 12 , further comprising a buffer region in which no convex portion is provided at a boundary between the first concave-convex portion and the second concave-convex portion.

15. A step or a slope is provided at a boundary between the first uneven portion and the second uneven portion, The detection set according to claim 12 , wherein an area of ​​the step or the slope on the first uneven portion side is higher than an area of ​​the step or the slope on the second uneven portion side.

16. The detection set according to claim 12 , further comprising a recessed region in which a recess is provided at a boundary between the first concave-convex portion and the second concave-convex portion.

17. The detection set described in claim 12, wherein, when the first uneven portion and the second uneven portion are adjacent to each other, a ratio (P1 / P2) of a pitch (P1) between the convex portions in the first uneven portion to a pitch (P2) between the convex portions in the second uneven portion is 1.1 or more and 5 or less.

18. An introduction part for introducing the solution into the flow channel is further provided, the solution introduced into the flow channel is made up of a plurality of types of solutions, The introduction part is provided at a plurality of locations corresponding to the plurality of types of solutions. The detection set according to claim 10.

19. The detection unit is provided with an electrode unit, The detection unit detects a reaction of the reaction solution to the antibody or the antigen based on a current flowing in the electrode unit. The detection set according to claim 10.

20. A detection device having a flow path provided on a substrate formed of resin for transporting a reaction liquid, a solid phase portion provided in the flow path in which an antibody or an antigen is solidified, a detection portion for detecting a reaction of the reaction liquid to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions formed integrally with the substrate in the region in which the flow path is provided; a solution supplying device that supplies a plurality of solutions including the reaction solution to the detection device; having The solution supply device includes: a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; a positioning unit that positions the communication unit at a predetermined position where the communication unit communicates with the solution storage unit; having The fine uneven structure is a first concave-convex portion in which the plurality of convex portions are relatively sparsely provided; a second concave-convex portion in which the plurality of convex portions are provided relatively densely; having A detection set, wherein the first uneven portion and the second uneven portion are provided closer to one end of the flow path than the solid phase portion.

21. A detection device having a flow path provided on a substrate formed of resin for transporting a reaction liquid, a solid phase portion provided in the flow path in which an antibody or an antigen is solidified, a detection portion for detecting a reaction of the reaction liquid to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions formed integrally with the substrate in the region in which the flow path is provided; a solution supplying device that supplies a plurality of solutions including the reaction solution to the detection device; having The solution supply device includes: a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; a positioning unit that positions the communication unit at a predetermined position where the communication unit communicates with the solution storage unit; having A detection set having a region in which the protrusions are arranged in a diamond lattice pattern.

22. A detection device having a flow path provided on a substrate formed of resin for transporting a reaction liquid, a solid phase portion provided in the flow path in which an antibody or an antigen is solidified, a detection portion for detecting a reaction of the reaction liquid to the antibody or the antigen, and a fine uneven structure having a plurality of convex portions formed integrally with the substrate in the region in which the flow path is provided; a solution supplying device that supplies a plurality of solutions including the reaction solution to the detection device; having The solution supply device includes: a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; a positioning unit that positions the communication unit at a predetermined position where the communication unit communicates with the solution storage unit; having A detection set having a region in which the protrusions are arranged in a regular lattice pattern.

23. A detection device having a flow path provided on a substrate formed of resin for transporting a reaction liquid, a solid phase portion provided in the flow path in which an antibody or an antigen is solidified, a detection portion for detecting a reaction of the reaction liquid with the antibody or the antigen, and a fine uneven structure having a plurality of convex portions formed integrally with the substrate in the region in which the flow path is provided; a solution supplying device that supplies a plurality of solutions including the reaction solution to the detection device; having The solution supply device includes: a solution unit having a plurality of solution storage portions for storing the plurality of solutions, a communication unit provided for each of the solution storage sections, the communication unit having a plurality of communication sections that communicate between the inside and outside of the solution storage section and supply the solution to the detection device; a positioning unit that positions the communication unit at a predetermined position where the communication unit communicates with the solution storage unit; having The detection set, wherein the protrusion is provided as a pyramid.

24. A detection set as described in Claim 20, wherein the first uneven portion is provided on one end side of the flow path relative to the second uneven portion.

25. A detection set as described in claim 20, having a buffer region at the boundary between the first uneven portion and the second uneven portion where no convex portion is provided.

26. A step or a slope is provided at a boundary between the first uneven portion and the second uneven portion, The detection set according to claim 20 , wherein an area of ​​the step or the slope on the first uneven portion side is higher than an area of ​​the step or the slope on the second uneven portion side.

27. ​​A detection set as described in claim 20, having a recessed area with a recess at the boundary between the first uneven portion and the second uneven portion.

28. A detection set as described in claim 20, wherein when the first uneven portion and the second uneven portion are adjacent to each other, the ratio (P1 / P2) of the pitch (P1) between the convex portions in the first uneven portion to the pitch (P2) between the convex portions in the second uneven portion is 1.1 or greater and 5 or less.

29. Further comprising an introduction portion for introducing the solution into the flow path, the solution introduced into the flow channel is made up of a plurality of types of solutions, The introduction part is provided at a plurality of locations corresponding to the plurality of types of solutions. The detection set according to claim 20.

30. The detection unit is provided with an electrode unit, The detection unit detects a reaction of the reaction solution to the antibody or the antigen based on a current flowing in the electrode unit. The detection set according to claim 20.

31. A detection method for detecting a reaction of a liquid sample with an antibody, using the detection set according to any one of claims 10 and 20 to 23.

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