Method of manufacturing immunochromatographic test kit

Abstract

To provide an immunochromatographic test kit manufacturing method which allows for more stably manufacturing immunochromatographic test kits compared with conventional manufacturing method.SOLUTION: A method of manufacturing an immunochromatographic test kit is provided, the method comprising preparing a carrier 6 consisting of porous member, and forming a plurality of detection sections by applying a capture antibody to the carrier 6 using an application needle 21 holding the capture antibody having a property to bind to a detection target at a tip thereof.SELECTED DRAWING: Figure 10

Description

[Technical Field]

[0001] The present invention relates to a method for producing an immunochromatographic test kit. [Background technology]

[0002] Immunochromatography is currently being implemented in society as a diagnostic aid for various diseases, primarily influenza viruses. Its principle is based on the antigen-antibody reaction, and it is widely used in medical settings due to its simplicity and effectiveness.

[0003] In conventional immunochromatography, the presence or absence of color development at the detection section of an immunochromatography test kit (also called a developing support or chromatographic medium) is visually observed. Specifically, when a target substance to be detected bound to a labeled antibody is captured by a capture antibody immobilized on the detection section and sufficiently accumulates on the detection section, color development resulting from the labeled antibody is visually confirmed.

[0004] International Publication No. 2010 / 061772 (Patent Document 1) discloses an immunochromatographic test kit in which a detection unit is formed with a size of several millimeters or more that allows visual determination. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2010 / 061772 Summary of the Invention [Problem to be solved by the invention]

[0006] Methods for forming the detection unit of the immunochromatographic test kit described above include the dispenser method and the inkjet method. In these methods, to form a detection unit with a small size of a few millimeters or less, the opening width of the nozzle tip must be made smaller.

[0007] However, when the liquid containing the capture antibody has high viscosity, clogging occurs when the opening width at the nozzle tip is reduced.

[0008] Therefore, depending on the viscosity of the liquid, it is difficult to reduce the size of the detection section, and it is not possible to reduce the amount of expensive capture antibody used, making it difficult to reduce costs.

[0009] The main object of the present invention is to provide a method for producing an immunochromatographic test kit that can reduce the amount of capture antibody used and produce a low-cost immunochromatographic test kit compared to conventional methods for producing immunochromatographic test kits. [Means for solving the problem]

[0010] The method for producing an immunochromatographic test kit according to the present invention comprises, on a porous member, a sample dropping section where a sample is dropped, a conjugate section to which a labeled antibody having the property of binding to a target substance in the sample is attached, and at least one detection section to which a capture antibody having the property of binding to the target substance is attached. The method for producing the immunochromatographic test kit comprises the step of applying a capture antibody using at least one applicator needle having the capture antibody at its tip to form at least one detection section.

[0011] In the above-mentioned method for producing an immunochromatographic test kit, the outer diameter of the tip of at least one applicator needle may be less than 1 mm.

[0012] In the method for producing the immunochromatographic test kit, in the step of forming and applying at least one detection portion, the capture antibody may be applied in the form of dots using at least one application needle.

[0013] In the manufacturing method of the above-mentioned immunochromatographic test kit, in the step of forming at least one detection portion, a recess may be formed in the porous member using at least one application needle, and at the same time, a capture antibody may be applied to the bottom surface of the recess.

[0014] In the manufacturing method of the above-mentioned immunochromatographic test kit, in the step of forming at least one detection part, multiple detection parts may be formed by applying the capture antibody one-dimensionally or two-dimensionally at intervals using multiple application needles.

[0015] In the method for manufacturing the immunochromatographic test kit, the plurality of application needles may include a first application needle carrying at its tip a first capture antibody that has the property of binding to a first type of detection target, and a second application needle carrying at its tip a second capture antibody that has the property of binding to a second type of detection target. In the step of forming the plurality of detection units, the first application needle may be used to apply the first capture antibody to a first region to form a first detection unit, and the second application needle may be used to apply the second capture antibody to a second region to form a second detection unit.

[0016] In the method for manufacturing the immunochromatographic test kit, the plurality of application needles may include a first application needle carrying at its tip a first capture antibody that has a property of binding to a first type of detection target, a second application needle carrying at its tip a second capture antibody that has a property of binding to a second type of detection target, and a third application needle carrying at its tip a third capture antibody that has a property of binding to a third type of detection target. In the step of forming the plurality of detection units, the first application needle may be used to apply the first capture antibody to a first region to form a first detection unit, the second application needle may be used to apply the second capture antibody to a second region to form a second detection unit, and the third application needle may be used to apply the third capture antibody to a third region to form a third detection unit.

[0017] In the above-described method for producing an immunochromatographic test kit, the sample dropping portion, the conjugate portion, and the at least one detection portion may be formed on a single porous member.

[0018] In the above-described method for producing an immunochromatographic test kit, the specimen dropping portion, the conjugate portion, and the at least one detection portion may be formed on a plurality of porous members. [Effects of the Invention]

[0019] According to the present invention, it is possible to provide a method for producing an immunochromatographic test kit that can reduce the amount of capture antibody used and produce a low-cost immunochromatographic test kit compared to conventional methods for producing immunochromatographic test kits. [Brief explanation of the drawings]

[0020] [Figure 1] FIG. 1 is a perspective view of an immunochromatographic test kit according to a first embodiment. [Figure 2] FIG. 2 is a partially enlarged plan view of the detection unit of the immunochromatography test kit shown in FIG. [Figure 3] FIG. 10 is a partial enlarged view for explaining the width of the detection portion. [Figure 4] FIG. 4 is a partially enlarged cross-sectional view taken along the arrows IV-IV in FIG. 2. [Figure 5] 3 is a flowchart of a method for producing an immunochromatographic test kit according to the first embodiment. [Figure 6] 1 is a perspective view of an application device used in the manufacturing method of the immunochromatographic test kit according to the first embodiment. FIG. [Figure 7] FIG. 7 is a front view of the application unit of the application device shown in FIG. 6. [Figure 8] FIG. 7 is a side view of the application unit of the application device shown in FIG. 6. [Figure 9] 7 is a front view illustrating the operation of the coating unit of the coating apparatus shown in FIG. 6. FIG. [Figure 10] 7 is a front view illustrating the operation of the coating unit of the coating apparatus shown in FIG. 6. FIG. [Figure 11] 3 is a partially enlarged plan view showing a modified example of the detection unit shown in FIG. 2. FIG. [Figure 12] 10 is a partially enlarged plan view showing another modified example of the detection unit shown in FIG. 2. FIG. [Figure 13] FIG. 10 is a partially enlarged plan view of a detection unit of the immunochromatographic test kit according to the second embodiment. [Figure 14] 14 is a partially enlarged plan view showing a modified example of the detection unit shown in FIG. 13. FIG. [Figure 15]14 is a partially enlarged plan view showing another modified example of the detection unit shown in FIG. 13. FIG. [Figure 16] 14 is a partially enlarged plan view showing still another modified example of the detection section shown in FIG. 13. FIG. [Figure 17] FIG. 10 is a perspective view of an immunochromatographic test kit according to a third embodiment. [Figure 18] FIG. 10 is a perspective view of an immunochromatographic test kit according to a fourth embodiment. [Figure 19] 1 is an electron microscope image obtained by observing the detection unit 3 of Example 1 using an electron microscope. [Figure 20] 10 is an electron microscope image obtained by observing the detection unit 3 of Example 2 using an electron microscope. [Figure 21] 10 is an electron microscope image obtained by observing the detection unit 3 using an electron microscope before the auxiliary liquid is dropped in Example 2. [Figure 22] 10 is an electron microscope image obtained by observing the detection unit 3 after the auxiliary liquid has been dropped using an electron microscope in Example 2. DETAILED DESCRIPTION OF THE INVENTION

[0021] Hereinafter, an embodiment of the present invention will be described as an example with reference to the drawings. In the following drawings, the same or corresponding parts are designated by the same reference numerals, and the description thereof will not be repeated.

[0022] (Embodiment 1) <Immunochromatography test kit> 1 and 2, the immunochromatographic test kit 100 according to the first embodiment mainly comprises a specimen dropping portion 1, a conjugate portion 2, a detection region t including a plurality of detection portions 3, a control region c including a control portion 4, and an absorption portion 5. As shown in FIGS. 1 and 2, the specimen dropping portion 1, the conjugate portion 2, the detection region t, the control region c, and the absorption portion 5 are arranged in the development direction D in the order described above.

[0023] The specimen is dropped onto the specimen dropping section 1. The specimen dropping section 1 is made of a porous material such as a glass fiber pad, a cellulose fiber pad, or a polyester pad.

[0024] The conjugate portion 2 is a portion to which a labeled antibody capable of binding to the target substance in the sample is immobilized. The labeled antibody is a conjugate of an antibody that specifically recognizes and binds to a first portion (first epitope) of the target substance and a labeling substance. The antibody can be selected arbitrarily depending on the target substance. The labeling substance can be selected arbitrarily from labeling substances used in conventional immunochromatography. The labeling substance includes, for example, at least one selected from the group consisting of metal nanoparticles (metal fine particles), latex fine particles, organic polymer fine particles, inorganic fine particles, and color-forming fine particles such as liposomes containing a coloring agent. The labeling substance includes, for example, at least one selected from the group consisting of noble metal nanoparticles such as gold nanoparticles, platinum nanoparticles, platinum-gold nanoparticles, and silver nanoparticles, titanium nanoparticles, iron nanoparticles, nickel nanoparticles, and cadmium nanoparticles. The metal nanoparticles may be colloidal metal nanoparticles with a particle size of 1 nm to 100 nm. The conjugate portion 2 is prepared, for example, by applying a suspension containing a labeled antibody to a porous member such as a glass fiber pad, a cellulose fiber pad, or a polyester pad, and then drying it.

[0025] The detection region t is disposed on the opposite side of the conjugate portion 2 from the specimen dropping portion 1 in the development direction D. The detection region t is formed on a carrier 6 made of a porous material. The detection region t includes a plurality of detection portions 3. Each detection portion 3 is connected to the specimen dropping portion 1 via the conjugate portion 2. Each detection portion 3 is disposed on the opposite side of the conjugate portion 2 from the specimen dropping portion 1 in the development direction D. Each detection portion 3 is a portion to which a capture antibody having the property of binding to the detection target is immobilized. The capture antibody is an antibody that specifically recognizes and binds to a second portion (second epitope) of the detection target that is different from the first portion. From a different perspective, the capture antibody has the property of binding to the test target that is bound to a labeled substance. The capture antibody can be selected arbitrarily depending on the detection target.

[0026] As shown in FIG. 2, the out-of-plane shape of each detection unit 3 is a dot. The dot width of each detection unit 3 is the smallest size that can be observed visually, or a size that cannot be observed visually but can be observed using a microscope. The dot width of each detection unit 3 is, for example, less than 1 mm. Preferably, the dot width of each detection unit 3 is equivalent to the field of view of an electron microscope or the like that can directly observe labeled substances of 1 μm or less in size that bind to labeled antibodies. The dot width of each detection unit 3 is, for example, 100 μm or less.

[0027] The width of the detection unit 3 is measured as follows. First, a sample containing a sufficient amount of the target substance is dropped onto the sample dropping unit 1 to induce an antigen-antibody reaction in the immunochromatographic test kit 100. Next, an electron microscope image of the detection unit 3 is acquired according to the test method described below. When the detection unit 3 includes the bottom surface 7A of the recess 7, as in the immunochromatographic test kit 100 according to the first embodiment, the electron microscope image acquired with the focus on the bottom surface 7A is used. Next, the outline of the area in which the labeled antibody is observed in the image is identified as the outline of the detection unit 3. Next, the width of the outline of the detection unit 3 is measured. If the outline of the area in which the labeled antibody is observed undulates inward and outward as shown in FIG. 3, the diameter D1 of the circumscribing circle CC of the outline and the diameter D2 of the inscribing circle IC of the outline are calculated by image processing, and the intermediate value between the diameters D1 and D2 is defined as the width of the outline of the detection unit 3.

[0028] 2 are arranged at intervals in a direction intersecting (e.g., perpendicular to) the deployment direction D. The detection units 3 are arranged one-dimensionally in a direction intersecting (e.g., perpendicular to) the deployment direction D. The distance between two adjacent detection units 3 is, for example, less than 1 mm.

[0029] The out-of-plane shape of each detection unit 3 is not particularly limited as long as it is a dot shape, but may be, for example, a circular shape. Note that the out-of-plane shape of each detection unit 3 may also be an ellipse, a square, a rectangle, or the like.

[0030] As shown in FIG. 4, multiple recesses 7 are formed in the detection region t of the carrier 6. Each recess 7 is recessed relative to the top surface of the carrier 6. Each recess 7 has a bottom surface 7A and a wall surface 7B connecting the bottom surface 7A to the top surface of the carrier 6. The opening of the top surface 6A and the bottom surface 7A are dot-shaped. In the cross section shown in FIG. 4, the bottom surface 7A and the wall surface 7B form an obtuse angle. In other words, the cross-sectional shape of each recess 7 shown in FIG. 4 has a tapered shape in which the distance between the opposing wall surfaces 7B narrows toward the bottom surface 7A. However, the diameter of the opening of the top surface 6A and the diameter of the bottom surface 7A may be the same. The width of the bottom surface 7A of each recess 7 is the minimum size that can be visually observed, for example, less than 1 mm. Preferably, the width of the bottom surface 7A of each recess 7 is equivalent to the field of view of an electron microscope or the like that allows direct observation of labeled substances of 1 μm or less in size that bind to labeled antibodies. For example, the dot width of each bottom surface 7A is 100 μm or less. The depth of each bottom surface 7A is greater than the focal depth of an electron microscope. The depth of each bottom surface 7A is, for example, 10 μm or more.

[0031] The out-of-plane shape of each bottom surface 7A is, for example, a circle, but the out-of-plane shape of each bottom surface 7A may also be an oval, square, rectangular, or the like.

[0032] Each detection unit 3 includes a bottom surface 7A of each recess 7. Each detection unit 3 is formed in each recess 7 of the carrier 6. Each detection unit 3 and each recess 7 are formed simultaneously, for example, on the carrier 6. A method for simultaneously forming the detection units 3 and the recesses 7 on the carrier 6 will be described in detail in the method for manufacturing an immunochromatography test kit described below.

[0033] The control region c is disposed on the opposite side of the detection region t from the conjugate region 2 in the development direction D. The control region C includes a control region 4. The control region 4 is a region where a control antibody that binds to the labeled antibody is immobilized. The out-of-plane shape of the control region 4 may be any shape, such as a line shape.

[0034] The control section 4 is formed on the carrier 6. The control section 4 is disposed on the opposite side of the detection section 3 from the conjugate section 2 in the development direction D.

[0035] For ease of explanation, the ranges of the detection region t and the control region c are indicated by dashed lines in Figures 1 and 2. In an actual immunochromatographic test kit, for example, the dashed lines are not shown.

[0036] The absorption section 5 is a section that absorbs excess sample and the like after development. The absorption section 5 is formed of a porous material such as a glass fiber pad, a cellulose fiber pad, or a polyester pad. The absorption section 5 is disposed on the opposite side of the control region c from the detection region t in the development direction D.

[0037] The out-of-plane shape of the carrier 6 is rectangular. The carrier 6 has a longitudinal direction aligned with the development direction D and a transverse direction perpendicular to the development direction D. The material constituting the carrier 6 may be any porous material, including, for example, at least one of nitrocellulose and polyvinylidene fluoride (PVDF). That is, in the immunochromatographic test kit 100 according to the first embodiment, the multiple detection sections 3 and control sections 4 are formed on porous members separate from the porous members constituting the specimen dropping section 1, the conjugate section 2, and the absorption section 5. The porous member constituting the specimen dropping section 1 is bonded to, for example, the porous member constituting the conjugate section 2. Each porous member constituting the conjugate section 2 and the absorption section 5 is bonded to, for example, each porous member constituting the carrier 6.

[0038] The specimen dropping section 1, the conjugate section 2, the detection section 3, the control section 4, the absorption section 5, and the carrier 6 may be fixed to a substrate (also called a backing sheet) (not shown) using an adhesive or an adhesive sheet. The carrier 6 may be a thin film formed on the substrate.

[0039] <Testing method using immunochromatographic test kit> In the testing method using the immunochromatographic test kit, the detection section 3 after the antigen-antibody reaction on the immunochromatographic test kit is observed using an electron microscope. The antigen-antibody reaction on the immunochromatographic test kit will be described below.

[0040] First, a sample is dropped onto sample dropping section 1. The dropped sample flows in the development direction D due to capillary action. The analyte in the sample binds to the labeled antibody in conjugate section 2. The conjugate of the analyte and labeled antibody moves in the development direction D along with the labeled antibody that is not bound to the analyte due to capillary action. The analyte that has bound to the labeled antibody binds to the capture antibody in detection section 3. As a result, the labeled antibody that has bound to the analyte accumulates in detection section 3. The labeled substance that has bound to the labeled antibody in detection section 3 is directly observed using an electron microscope. Furthermore, the labeled antibody that has not bound to the analyte binds to the control antibody in control section 4. As a result, the labeled antibody that has not bound to the analyte is captured in control section 4. The labeled substance that has bound to the labeled antibody in control section 4 is directly observed using an electron microscope. Excess sample that flows downstream of control section 4 in the development direction D is absorbed by absorption section 5.

[0041] The test method using the immunochromatographic test kit is carried out, for example, as follows. First, a specimen is dropped into specimen dropping section 1, and after a specified time has passed, an auxiliary liquid is dropped.

[0042] The auxiliary liquid has conductivity to prevent static electricity and heat generation, which contribute to image clarity under electron microscope measurement conditions, and / or the ability to polymerize to form a film. The auxiliary liquid contains, as essential components, at least one compound selected from glycerin and glycerin substitutes; polysorbates such as polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, and polysorbate 85; and polysorbate substitutes, and optionally contains at least one compound selected from monosaccharides, disaccharides, salts, and buffer solutions.

[0043] Next, the detection unit 3 is observed using an electron microscope. Specifically, first, the carrier 6 is set in the chamber of the electron microscope. Next, the detection unit 3 is searched for while observing the top surface of the carrier 6. In the immunochromatography test kit 100, defocusing due to the distance between the top surface of the carrier 6 and the bottom surface 7A is observed in the microscopic image (bright-field image) of the electron microscope. Therefore, by checking whether or not there is defocusing, the recess 7 can be searched for in the microscopic image, and the detection unit 3 can be easily searched for in the image. Thereafter, the image can be focused on the bottom surface 7A, thereby focusing the image on the detection unit 3. Next, an electron microscope image is captured, and the number of labeled substances in the image is measured. Since the outlines of the labeled substances can be clearly identified in the electron microscope image, the number of labeled substances in the image can be counted. If the measured number of labeled substances exceeds a predetermined criterion, it is determined that the substance to be detected is present in the sample (i.e., "positive").

[0044] The above observation and measurement are performed in the same manner for, for example, a plurality of detection units 3. In this case, for example, the average, maximum, minimum, etc. of the number of labeled substances in the same field of view of each detection unit 3 is calculated, and any one of these calculated values ​​is used as the evaluation value, and if the evaluation value exceeds a predetermined judgment criterion, it is judged as "positive."

[0045] <Manufacturing method of immunochromatography test kit> Next, we will explain an example of a method for manufacturing the immunochromatographic test kit 100. As shown in Fig. 5, the method for manufacturing the immunochromatographic test kit 100 includes a step (S1) of preparing a carrier 6 and a step (S2) of forming a plurality of detection units 3 on the prepared carrier 6.

[0046] In the step of preparing the carrier 6, a carrier 6 having the control section 4 formed on the upper surface thereof, or a carrier 6 having no control section 4 formed on the upper surface thereof is prepared.

[0047] In the process of forming the multiple detection portions 3, a coating needle 21 (see FIG. 10) holding a capture antibody at its tip is used to coat the upper surface of the carrier 6 with the capture antibody in the form of dots. The diameter of the tip of the coating needle 21 (hereinafter referred to as the tip diameter) is less than 1 mm. In this process, the coating needle 21 is used to form recesses 7 in the carrier 6, and at the same time, the capture antibody is coated onto the bottom surface 7A of the recesses 7. In other words, the coating needle 21 holding the capture antibody at its tip is pressed further into the carrier 6 after coming into contact with the upper surface of the carrier 6. In this process, a coating device shown in FIGS. 6 to 10 is used. The configuration of the coating device will be described later.

[0048] In this way, a carrier 6 including a plurality of detection units 3 is formed. If the control unit 4 is formed on this carrier 6, the conjugate unit 2 and the absorption unit 5 are connected to the carrier 6, and the specimen dropping unit 1 is further connected to the conjugate unit 2, thereby producing an immunochromatographic test kit 100. If the control unit 4 is not formed on the carrier 6, the control unit 4 is formed on the carrier 6. Thereafter, the conjugate unit 2 and the absorption unit 5 are connected to the carrier, and the specimen dropping unit 1 is further connected to the conjugate unit 2, thereby producing an immunochromatographic test kit 100. In the method for producing the immunochromatographic test kit 100, the specimen dropping unit 1, the conjugate unit 2, the control unit 4, and the absorption unit 5 can each be formed in the same manner as those in conventional immunochromatographic test kits.

[0049] <Configuration of coating equipment> Next, an example of an application device used to form a plurality of detection sections 3 in the manufacturing method of the immunochromatographic test kit 100 will be described.

[0050] 6, the coating apparatus according to the first embodiment mainly includes an X-axis table 11, a Y-axis table 12, a Z-axis table 13, a coating mechanism 14, an observation optical system 15, a CCD camera 16, and a control unit. The X-axis table 11, the Y-axis table 12, the Z-axis table 13, the coating mechanism 14, the observation optical system 15, and the CCD camera 16 are arranged, for example, inside a processing chamber. The control unit includes an operation panel 17, a monitor 18, and a control computer 19. The operation panel 17, the monitor 18, and the control computer 19 are arranged, for example, outside the processing chamber.

[0051] The Y-axis table 12 is movable in the Y-axis direction and is capable of carrying the carrier 6. For example, a guide rail extending along the Y-axis direction is fixed to the bottom of the processing chamber. A guide unit movable along the guide rail is connected to the underside of the Y-axis table 12. The upper surface of the Y-axis table 12 serves as a mounting surface on which the carrier 6 is mounted. A structure spanning the Y-axis table 12 in the X-axis direction is fixed to the bottom of the processing chamber.

[0052] The X-axis table 11 is disposed on a structure that is installed so as to straddle the Y-axis table 12 in the X-axis direction. A movable body, to which the Z-axis table 13 is connected, is installed on the X-axis table 11 so as to be movable in the X-axis direction. The movable body is movable in the X-axis direction, for example, using a ball screw. The X-axis table 11 is fixed to the bottom surface of the processing chamber via the structure. Therefore, the Y-axis table 12 described above is movable in the Y-axis direction relative to the X-axis table 11.

[0053] As described above, the Z-axis table 13 is installed on a movable body connected to the X-axis table 11. The observation optical system 15 and the coating mechanism 14 are connected to the Z-axis table 13. The observation optical system 15 is used to observe the coating position on the carrier 6. The CCD camera converts the observed image into an electrical signal. The Z-axis table 13 holds the observation optical system 15 and the coating mechanism 14 so that they can move in the Z-axis direction.

[0054] The operation panel 17, monitor 18, and control computer 19 are used to control the Y-axis table 12, X-axis table 11, Z-axis table 13, observation optical system 15, and coating mechanism 14. The operation panel 17 is used to input commands to the control computer 19. The monitor 18 displays the image data converted by the above-mentioned CCD camera 16 and the output data from the control computer 19.

[0055] <Configuration of Coating Mechanism 14> Next, the detailed configuration of the coating mechanism 14 shown in Fig. 6 will be described. As shown in Figs. 7 and 8, the coating mechanism 14 mainly includes a servo motor 41, a base 42, a cam 43, a bearing 44, a cam connecting plate 45, a movable part 46, a coating needle holder 20, a coating needle 21 held by the coating needle holder 20, and a coating material container 22. The servo motor 41 is installed so that its rotation axis extends in the Z-axis direction shown in Fig. 6. The servo motor 41 is held by the base 42. A cam 43 is connected to the rotation axis of the servo motor 41. The cam 43 is rotatable around the rotation axis of the servo motor 41.

[0056] The cam 43 includes a central portion connected to the rotary shaft of the servo motor 41 and a flange portion connected to one end of the central portion. The upper surface of the flange portion (the surface facing the servo motor 41) forms a cam surface. This cam surface is formed in an annular shape along the outer periphery of the central portion and is formed in a sloped shape so that the distance from the bottom surface of the flange portion varies. Specifically, the cam surface includes an upper end flat region that is farthest from the bottom surface of the flange portion, a lower end flat region that is shortest from the bottom surface of the flange portion, and a sloped region connecting the upper end flat region and the lower end flat region. The upper end flat region is spaced apart from the lower end flat region in the circumferential direction relative to the rotary shaft.

[0057] The bearing 44 has an outer ring arranged to contact the cam surface of the cam 43, and an inner ring connected to the cam connecting plate 45. As shown in FIG. 7, the bearing 44 is arranged in a specific direction (to the right of the servo motor 41) when viewed from the cam 43. The bearing 44 is biased toward the cam surface of the cam 43 by a spring 50, which will be described later. Therefore, when the cam 43 rotates, the outer peripheral surface of the outer ring of the bearing 44 is held in contact with the cam surface. The cam connecting plate 45 has one end connected to the inner ring of the bearing 44 and the other end fixed to the movable part 46. A coating needle holder fixing part 47 and a coating needle holder storage part 48 are connected to the movable part 46.

[0058] A fixed pin 51 is installed on the base 42. A fixed pin 52 is installed on the movable part 46. One end of a spring 50 is connected to the fixed pin 51, and the other end of the spring 50 is connected to the fixed pin 52. The movable part 46 is subjected to a tensile force toward the coating material container 22 by the spring 50. The tensile force of this spring 50 acts on the bearing 44 via the movable part 46 and the cam connecting plate 45. The tensile force of this spring 50 keeps the bearing 44 pressed against the cam surface of the cam 43.

[0059] The movable part 46, the coating needle holder fixing part 47, and the coating needle holder storage part 48 are connected to a linear guide 49 installed on the stand 42. The linear guide 49 is arranged to extend in the Z-axis direction. Therefore, the movable part 46, the coating needle holder fixing part 47, and the coating needle holder storage part 48 are movable along the Z-axis direction.

[0060] The coating needle holder 20 is stored in a coating needle holder storage section 48. The coating needle holder 20 includes a coating needle 21. The coating needle 21 is arranged on the underside of the coating needle holder 20 so as to protrude from the coating needle holder 20. The extension direction of the coating needle 21 is along the direction of gravity (the Z-axis direction in FIG. 6). The shape of the tip of the coating needle 21 is, for example, a truncated cone. The tip diameter in the radial direction relative to the central axis of the coating needle 21 is less than 1 mm. Preferably, the tip diameter of the coating needle 21 is 100 μm or less.

[0061] A coating material container 22 is disposed below the coating needle holder 20. A space for storing a coating material 70 is formed inside the coating material container 22. As shown in FIGS. 9 and 10, a first hole and a second hole connecting the space to the outside are formed in the bottom and top of the coating material container 22. The first hole and the second hole may have any shape, but may be circular, for example. The first hole and the second hole are arranged so as to overlap in the extension direction of the coating needle 21. The coating material 70 contains a capture antibody.

[0062] As the coating needle holder storage section 48 moves in the Z-axis direction, the coating needle holder 20 and the coating needle 21 move up and down relative to the coating material container 22. This allows switching between a first state in which the tip of the coating needle 21 is immersed in the coating material 70 stored in the coating material container 22 and a second state in which the tip protrudes downward from the bottom surface of the coating material container 22 through a hole formed in the bottom of the coating material container 22.

[0063] <Operation of the coating device> Next, the operation of the coating device in the step of forming multiple detection units 3 in the manufacturing method of the immunochromatographic test kit will be described. First, the carrier 6 is loaded onto the Y-axis table 12. Next, the tip of the coating needle 21 is immersed in the coating material 70 stored in the coating material container 22, thereby placing the carrier 6 in the first state. Next, the X-axis table 11 and the Y-axis table 12 are operated to position the carrier 6 in the X-axis and Y-axis directions so that the area where the detection units 3 are to be formed is located directly below the coating mechanism 14. Next, the Z-axis table 13 is operated to lower the Z-axis table 13 and the coating mechanism 14 by a distance d in the Z-axis direction so that the tip of the coating needle 21 is pushed below the top surface of the carrier 6 when the coating needle is placed in the second state.

[0064] Next, the servo motor 41 of the coating mechanism 14 is operated to switch from the first state to the second state. Specifically, operating the servo motor 41 rotates the rotation shaft of the servo motor 41, thereby rotating the cam 43. As a result, the height of the cam surface of the cam 43 changes in the Z-axis direction, and the position of the bearing 44 in contact with the cam surface also changes in the Z-axis direction in response to the rotation of the drive shaft of the servo motor 41. In response to the positional change of the bearing 44 in the Z-axis direction, the movable part 46, the coating needle holder fixing part 47, the coating needle holder storage part 48, and the coating needle holder 20 held in the coating needle holder storage part 48 also move in the Z-axis direction. As a result, operating the servo motor 41 moves the coating needle 21 in the Z-axis direction, switching between the first state shown in FIG. 9 and the second state shown in FIG. 10. In the coating mechanism 14, the rotational movement of the servo motor 41 can be converted into the movement (up and down movement) of the coating needle 21 in the Z-axis direction, so that the coating needle 21 can be moved quickly and accurately in the Z-axis direction.

[0065] 9, application needle 21 is positioned at the uppermost position in its movable range. At this time, the tip of application needle 21 is immersed in application material 70 held in application material container 22. In the first state, the outer ring of bearing 44 is in contact with the upper flat region of the cam surface of cam 43.

[0066] 10, the coating needle 21 is positioned at the lowest position in its movable range. At this time, the tip of the coating needle 21 holds a coating material containing a capture antibody. Specifically, the coating material containing a capture antibody is held on the end face and side surface of the tip of the coating needle 21. The tip of the coating needle 21 passes through a hole formed in the bottom of the coating material container 22, protrudes downward from the bottom surface of the coating material container 22, and is pushed below the upper surface of the carrier 6.

[0067] In this way, by operating the servo motor 41 to switch from the first state to the second state, recesses 7 are formed in the carrier 6 and the capture antibody is simultaneously applied to the bottom surfaces 7A of the recesses 7. Next, the servo motor 41 is operated to switch from the second state to the first state.

[0068] Furthermore, only the X-axis table 11 and the Y-axis table 12 are operated to position the area where the second detection unit 3 is to be formed directly under the coating mechanism 14, and then the Z-axis table 13 is lowered by a distance d, and then the servo motor 41 of the coating mechanism 14 is operated to switch from the first state to the second state. Furthermore, after switching from the second state to the first state, the Z-axis table 13 is raised by a distance d.

[0069] By continuously repeating the above operation, a plurality of detection portions 3 are formed on the upper surface of the carrier 6. <Action and effect> The effects of the manufacturing method of the immunochromatographic test kit 100 according to embodiment 1 will be explained in comparison with a manufacturing method in which each detection section 3 is formed using a dispenser (hereinafter simply referred to as Comparative Example 1), and a manufacturing method in which each detection section 3 is formed by an inkjet method (hereinafter simply referred to as Comparative Example 2).

[0070] In Comparative Examples 1 and 2, in order to form the detection unit 3 in a minute size, it is necessary to reduce the opening width at the tip of the dispenser or nozzle. However, when a highly viscous coating material 70 is used, the coating material 70 cannot be ejected, and clogging may occur. Furthermore, in Comparative Example 2, in a method in which a piezoelectric element or the like is used to instantaneously increase the pressure inside a container to eject a liquid material containing a capture antibody, the ejection pressure is high, so the liquid material tends to spread when it hits the carrier 6, resulting in problems such as an unstable shape of the detection unit 3.

[0071] In contrast, in the manufacturing method of the immunochromatographic test kit 100, the tip diameter of the applicator needle 21 can be reduced to form a minute detection unit 3, and therefore, regardless of the viscosity, clogging does not occur as in Comparative Examples 1 and 2. Furthermore, in the above manufacturing method, the position of the tip of the applicator needle 21 in the Z-axis direction relative to the upper surface of the carrier 6 can be precisely controlled by the Z-axis table 13, so that variation in the size of the detection unit 3 can be reduced and the immunochromatographic test kit 100 can be stably manufactured.

[0072] Furthermore, in Comparative Example 1, since droplets are applied, it is difficult to stably form a detection portion 3 with a dot width of less than 1 mm, and in particular to stably form a detection portion 3 with a dot width of less than 100 μm.

[0073] In contrast, the manufacturing method of the immunochromatographic test kit 100 uses an application needle 21, so that the immunochromatographic test kit 100 can be easily and stably manufactured in which the out-of-plane shape of each detection part 3 is dot-shaped and the width of each detection part 3 is less than 1 mm.

[0074] Furthermore, in the manufacturing method of the immunochromatography test kit 100, the recesses 7 are formed in the carrier 6 using the application needle 21, and at the same time, the capture antibody is applied to the bottom surface 7A of the recesses 7. Therefore, the formation of the detection area 3 including the bottom surface 7A of one recess 7 and the application of the capture antibody can be performed in a single operation. Therefore, there is no increase in takt time due to the formation of the detection area 3, and there is no increase in manufacturing costs.

[0075] Next, the effects of the immunochromatographic test kit 100 according to the first embodiment will be explained in comparison with a conventional immunochromatographic test kit (hereinafter simply referred to as Comparative Example 3). In Comparative Example 1, the detection part is formed on a flat surface, and is provided so that the color development at the detection part resulting from the labeled antibody can be visually confirmed. Therefore, in Comparative Example 1, the area of ​​the detection part is set to be sufficient (for example, several mm) so as to prevent erroneous determination by visual observation. 2 As a result, it was difficult to reduce the amount of expensive capture antibody immobilized on the detection part. As a result, it was difficult to reduce the production cost in Comparative Example 1.

[0076] In contrast, in the immunochromatographic test kit 100, the dot width of each detection unit 3 can be made less than 1 mm by reducing the tip diameter of the applicator needle 21. In other words, in the immunochromatographic test kit 100, the area of ​​each detection unit 3 can be made smaller than the area of ​​each detection unit in Comparative Example 3, and therefore the amount of capture antibody immobilized on each detection unit 3 can be reduced compared to the amount of capture antibody immobilized on each detection unit in Comparative Example 3. As a result, the manufacturing cost of the immunochromatographic test kit 100 can be reduced compared to the manufacturing cost of Comparative Example 3.

[0077] Furthermore, according to the immunochromatographic test kit 100, the detection unit 3 can be observed using an electron microscope, and the number of labeled antibodies captured by the detection unit 3 in the electron microscope image can be measured, thereby quantitatively evaluating the substance to be detected in the sample.

[0078] Furthermore, when the detection unit formed on a flat surface is miniaturized as in Comparative Example 1, it is necessary to magnify the detection unit using a microscope or the like to confirm the labeled antibody, but it is difficult to locate and align the tiny detection unit using a microscope. One possible solution to this problem is to provide a marker on the kit and then provide the detection unit at a predetermined distance from that marker. However, this requires separate equipment for providing the marker, and the number of processes increases, making it difficult to reduce the manufacturing takt time and making it difficult to reduce costs.

[0079] In contrast, in the immunochromatographic test kit 100, a plurality of recesses 7 are formed in the carrier 6, and each detection unit 3 includes the bottom surface 7A of one recess 7. Therefore, when the labeled substance on the detection unit 3 is directly observed using an electron microscope, the defocus caused by the distance between the top surface and the bottom surface 7A of the carrier 6 can be confirmed in the microscopic image of the electron microscope, and this defocus can be used to search for the recesses 7 in the microscopic image of the electron microscope. As a result, in the immunochromatographic test kit 100, each detection unit 3 can be made smaller than in Comparative Example 1.

[0080] Furthermore, by focusing the image on the bottom surface 7A, the image can be easily focused on the detection unit 3. That is, in the immunochromatography test kit 100, the time required to search for the detection unit 3 can be shortened compared to when each detection unit 3 does not include the bottom surface 7A of one recess 7.

[0081] Furthermore, in the immunochromatographic test kit 100, unlike Comparative Example 1, there is no need to provide a separate mark to be used when searching for the detection unit 3, and there is no need to prepare equipment for providing the mark separately from equipment for forming the detection unit. Furthermore, in the immunochromatographic test kit 100, the recess 7 can be formed simultaneously with the detection unit 3, so the manufacturing takt time can be reduced compared to when a mark is provided as in Comparative Example 1.

[0082] In the immunochromatographic test kit 100, the width of each detection part can be made as small as 100 μm or less. In such an immunochromatographic test kit 100, the amount of capture antibody immobilized on each detection part 3 is significantly reduced compared to the amount of capture antibody immobilized on each detection part in Comparative Example 3, and therefore the manufacturing cost of the immunochromatographic test kit 100 can be significantly reduced compared to the manufacturing cost of Comparative Example 3.

[0083] The immunochromatographic test kit 100 includes a plurality of detection units 3. In this immunochromatographic test kit 100, the labeling substances of the plurality of detection units 3 can be confirmed in the electron microscope image, compared to an immunochromatographic test kit that includes only one detection unit 3, and therefore the reliability of the test results is improved.

[0084] In the immunochromatographic test kit 100, the multiple detection units 3 are arranged one-dimensionally in the development direction D. Compared to an immunochromatographic test kit in which the multiple detection units 3 are arranged randomly, in this immunochromatographic test kit 100, the detection target efficiently flows to all of the detection units 3 and binds to the capture antibody, thereby obtaining stable test results.

[0085] In the immunochromatographic test kit 100, the shortest distance between two adjacent detection units 3 is less than 1 mm. In such an immunochromatographic test kit 100, the multiple detection units 3 can be observed even within a relatively small field of view, compared to immunochromatographic test kits in which the shortest distance is 1 mm or more, and therefore each detection unit 3 can be easily searched for.

[0086] <Variation 1> The immunochromatographic test kit 100 according to the first embodiment can be modified as follows.

[0087] The detection area t only needs to include at least one detection unit 3 . In the immunochromatographic test kit 100, the detection units 3 have the same configuration, but this is not limited to this. In the immunochromatographic test kit 100, the concentration of the capture antibody immobilized in each detection unit 3 is constant, but the concentration of the capture antibody immobilized in each detection unit 3 may be different from each other. For example, the detection units 3 may be configured so that the concentration of the capture antibody gradually decreases from one side to the other in a direction perpendicular to the development direction D. Also, a set of patterns consisting of multiple detection units 3 with different capture antibody concentrations may be arranged side by side. Such multiple detection units 3 can be easily formed by using multiple application mechanisms 14 with different capture antibody concentrations in the application material stored in the application material container 22.

[0088] Furthermore, at least one of the out-of-plane shape of each detection unit 3, the dot width of each detection unit 3, the distance between two adjacent detection units 3, the out-of-plane shape of the bottom surface 7A of each recess 7, the maximum width of each bottom surface 7A, and the depth of each bottom surface 7A may be different. Such multiple detection units 3 can be easily formed by using multiple types of application needles 21 or multiple types of application conditions.

[0089] In the immunochromatography test kit 100, a plurality of recesses 7 are formed in the detection region t of the carrier 6, and one detection part 3 includes the bottom surface 7A of one recess 7, but this is not limited to this.

[0090] Each detection unit 3 may include the entire bottom surface 7A and at least a part of the wall surface 7B of each recess 7. Alternatively, each detection unit 3 may include the entire bottom surface 7A and wall surface 7B of each recess 7. Alternatively, each detection unit 3 may include the entire bottom surface 7A and wall surface 7B of each recess 7, as well as an annular region of the upper surface of the carrier 6 that is continuous with the wall surface 7B.

[0091] The multiple detection units 3 may include at least one detection unit 3 shown in FIG. 4 and at least one detection unit 3 shown in FIG. 11. The multiple detection units 3 shown in FIG. 11 include only the upper surface of the carrier 6. Such detection units 3 can be easily formed in the above-described manufacturing method by causing the tip of the application needle 21 or the coating material 70 attached to the tip to come into contact with the upper surface of the carrier 6 and then move upward without being pushed downward. Furthermore, all of the detection units 3 may be configured as the detection units 3 shown in FIG. 11. In other words, the detection region t of the carrier 6 does not need to have multiple recesses 7 formed therein.

[0092] 12, in the immunochromatographic test kit 100, multiple detection units 3 may be arranged at intervals in the development direction D. The distance between two adjacent detection units 3 in the development direction D is equal to the distance between two adjacent detection units 3 in a direction perpendicular to the development direction D, for example.

[0093] In the manufacturing method of the immunochromatography test kit 100, one application needle 21 is used to form multiple detection units 3, but multiple application needles 21 may be used to form multiple detection units 3. In other words, the application device may be equipped with multiple application mechanisms 14.

[0094] Furthermore, to further improve the accuracy of the coating process of coating material 70 by coating needle 21, the movement speed of coating needle 21 in the Z-axis direction may be changed depending on the position of coating needle 21. For example, when switching from the first state to the second state, the movement speed of coating needle 21 is reduced just before the second state is achieved. The reduction in the movement speed of coating needle 21 is achieved by reducing the rotation speed of servo motor 41.

[0095] By performing such control, the moving speed of the coating needle 21 is sufficiently slow when the coating needle 21 comes into contact with the carrier 6, thereby lengthening the contact time between the tip of the coating needle 21 or the coating material 70 attached to the tip and the carrier 6. This increases the amount of capture antibody applied to the carrier 6 and increases the concentration, making it possible to further stabilize the antigen-antibody reaction with the detection target.

[0096] In the immunochromatographic test kit 100, the specimen dropping section 1 and the conjugate section 2 may be formed on a single porous member that is separate from the carrier 6. In the immunochromatographic test kit 100, the conjugate section 2 may be formed on the carrier 6, and the specimen dropping section 1 may be formed on a porous member that is separate from the carrier 6. In the immunochromatographic test kit 100, the absorption section 5 may be formed on the carrier 6.

[0097] (Embodiment 2) The immunochromatographic test kit 101 according to embodiment 2 has basically the same configuration and produces the same effects as the immunochromatographic test kit 100 according to embodiment 1, but differs from the immunochromatographic test kit 100 in that the multiple detection units 3 include a first detection unit 3A and a second detection unit 3B, as shown in FIG. 13.

[0098] A first capture antibody capable of binding to a first type of target substance is immobilized on the first detection section 3A, and a second capture antibody capable of binding to a second type of target substance that is different from the first type of target substance is immobilized on the second detection section 3B.

[0099] 13, the multiple detectors 3 include, for example, multiple first detectors 3A and multiple second detectors 3B. The multiple first detectors 3A are arranged at intervals from one another in a direction perpendicular to the deployment direction D. The multiple first detectors 3A are, for example, arranged one-dimensionally. The multiple second detectors 3B are, for example, arranged one-dimensionally, and the multiple second detectors 3B are, for example, arranged one-dimensionally.

[0100] The multiple first detection units 3A and multiple second detection units 3B are arranged at intervals in a direction perpendicular to the deployment direction D. The shortest distance between the first detection unit 3A and the second detection unit 3B is longer than the shortest distance between two adjacent first detection units 3A and the shortest distance between two adjacent second detection units 3B. The multiple first detection units 3A and multiple second detection units 3B are, for example, arranged one-dimensionally as a whole.

[0101] Each of the first detection unit 3A and the second detection unit 3B has the same configuration as each detection unit 3 in embodiment 1. The dot width of the first detection unit 3A is equal to the dot width of the second detection unit 3B, for example. Note that the dot width of the first detection unit 3A may be narrower or wider than the dot width of the second detection unit 3B, for example.

[0102] The detection area t is divided into a first detection area t1 where a plurality of first detection units 3A are arranged and a second detection area t2 where a plurality of second detection units 3B are arranged. The first detection area t1 and the second detection area t2 are arranged side by side in a direction perpendicular to the deployment direction D.

[0103] Each first detection portion 3A includes a bottom surface 7A of each recess 7. Each second detection portion 3B includes a bottom surface 7A of each recess 7.

[0104] According to the immunochromatographic test kit 101 of the second embodiment, two types of detection targets can be detected from a single specimen by immunochromatography.

[0105] The immunochromatographic test kit 101 can be manufactured in the same manner as the immunochromatographic test kit according to the first embodiment. Preferably, the coating device includes a first coating mechanism 14 for forming a first detection unit 3A by coating a first capture antibody on the carrier 6, and a second coating mechanism 14 for forming a second detection unit 3B by coating a second capture antibody on the carrier 6. The first coating mechanism 14 includes a first coating needle 21 and a first coating material container 22 in which a coating material containing the first capture antibody is stored. The second coating mechanism 14 includes a second coating needle 21 and a second coating material container 22 in which a coating material containing the second capture antibody is stored.

[0106] In the manufacturing method of the immunochromatographic test kit 101, the first application needle 21 holds a first capture antibody at its tip. The second application needle 21 holds a second capture antibody at its tip. In the step of forming the multiple detection parts 3, the first application needle 21 is used to apply the first capture antibody to a first region of the carrier 6, thereby forming a first detection part 3A. Furthermore, the second application needle 21 is used to apply the second capture antibody to a second region of the carrier 6, thereby forming a second detection part 3B.

[0107] By manufacturing the immunochromatographic test kit 101 using such an application device, the manufacturing labor hours can be reduced compared to manufacturing the immunochromatographic test kit 101 using a single application device equipped with only one application mechanism 14, and the manufacturing costs can be reduced compared to manufacturing the immunochromatographic test kit 101 using multiple application devices equipped with only one application mechanism 14.

[0108] <Variation 2> The immunochromatographic test kit 101 according to the second embodiment can also be modified in the same manner as the immunochromatographic test kit 100 according to the first embodiment.

[0109] Furthermore, the immunochromatographic test kit 101 can take on the following modifications. The plurality of first detection units 3A and the plurality of second detection units 3B may be arranged alternately.

[0110] 14 to 16, the multiple detection units 3 may further include multiple third detection units 3C in addition to the first detection unit 3A and the second detection unit 3B. A third capture antibody having the property of binding to a third type of detection target different from the first type of detection target and the second type of detection target is immobilized on each third detection unit 3C.

[0111] As shown in Figure 14, each of the multiple first detection units 3A, multiple second detection units 3B, and multiple third detection units 3C is arranged alternately, for example, one by one, in a direction perpendicular to the deployment direction D.

[0112] 15, the plurality of first detection units 3A, the plurality of second detection units 3B, and the plurality of third detection units 3C are arranged alternately in a direction perpendicular to the deployment direction D. In this case, the distance between two adjacent first detection units 3A and the distance between two adjacent second detection units 3B are shorter than the distance between adjacent first detection units 3A and second detection units 3B, for example. The distance between two adjacent second detection units 3B and the distance between two adjacent third detection units 3C are shorter than the distance between adjacent second detection units 3B and third detection units 3C, for example.

[0113] As shown in FIG. 16 , the plurality of first detection units 3A, the plurality of second detection units 3B, and the plurality of third detection units 3C are arranged one-dimensionally in, for example, the deployment direction D. Furthermore, the plurality of first detection units 3A, the plurality of second detection units 3B, and the plurality of third detection units 3C are arranged alternately one by one in a direction perpendicular to the deployment direction D. The distance between adjacent first detection units 3A and second detection units 3B in the direction perpendicular to the deployment direction D is longer than, for example, the distance between two adjacent first detection units 3A and two adjacent second detection units 3B in the deployment direction D. The distance between adjacent second detection units 3B and third detection units 3C in the direction perpendicular to the deployment direction D is longer than, for example, the distance between two adjacent second detection units 3B and two adjacent third detection units 3C in the deployment direction D.

[0114] In the modified examples shown in Figures 14 to 16, in a set of first detection unit 3A, second detection unit 3B, and third detection unit 3C, at least one of the out-of-plane shape of each detection unit 3, the dot width of each detection unit 3, the distance between two adjacent detection units 3, the out-of-plane shape of the bottom surface 7A of each recess 7, the maximum width of each bottom surface 7A, and the depth of each bottom surface 7A may be different from each other.

[0115] For example, the dot width of the second detection unit 3B may be narrower than the dot width of the first detection unit 3A and wider than the dot width of the third detection unit 3C. The maximum width of the bottom surface 7A included in the second detection unit 3B may be narrower than the maximum width of the bottom surface 7A included in the first detection unit 3A and wider than the maximum width of the bottom surface 7A included in the third detection unit 3C.

[0116] The second detection unit 3B may have a rectangular out-of-plane shape, and the first detection unit 3A and the third detection unit 3C may have a circular out-of-plane shape.

[0117] The depth of the bottom surface 7A included in the second detection portion 3B may be shallower than the maximum width of the bottom surface 7A included in the first detection portion 3A and deeper than the maximum width of the bottom surface 7A included in the third detection portion 3C.

[0118] The distance between the first detection unit 3A and the second detection unit 3B may be longer than the distance between the second detection unit 3B and the third detection unit 3C and the distance between the third detection unit 3C and the first detection unit 3A.

[0119] Such first detection unit 3A, second detection unit 3B, and third detection unit 3C can be easily formed by using a plurality of types of application needles 21 or by using a plurality of types of application conditions.

[0120] In this way, the first detection portion 3A, the second detection portion 3B, and the third detection portion 3C can be easily distinguished within one field of view.

[0121] (Embodiment 3) The immunochromatographic test kit 102 according to the third embodiment has basically the same configuration and produces the same effects as the immunochromatographic test kit 100 according to the first embodiment, but differs from the immunochromatographic test kit 100 in that the specimen dropping portion 1 and the conjugate portion 2 are formed on a carrier 6. That is, in the immunochromatographic test kit 102, the specimen dropping portion 1, the conjugate portion 2, the plurality of detection portions 3, and the control portion 4 are formed on the upper surface of a single porous member (one carrier 6).

[0122] In the immunochromatographic test kit 102 shown in FIG. 17, the absorbing part 5 is also formed on the carrier 6.

[0123] The specimen dropping section 1 is an area on the carrier 6 where the specimen is dropped, and is located on one side in the developing direction D. The conjugate section 2 is an area on the upper surface of the carrier 6 where a labeled antibody is applied, and is located between the specimen dropping section 1 and the detection area t in the developing direction D. The absorption section 5 is an area on the carrier 6 where excess specimen is absorbed, and is located on the other side of the control area C in the developing direction D.

[0124] According to immunochromatographic test kit 102 , the number of parts is reduced compared to immunochromatographic test kit 100 , and therefore the manufacturing costs of immunochromatographic test kit 102 can be reduced compared to the manufacturing costs of immunochromatographic test kit 100 .

[0125] <Variation 3> The immunochromatographic test kit 102 according to the third embodiment can also adopt the same modifications as the immunochromatographic test kit 100 according to the first embodiment and the immunochromatographic test kit 101 according to the second embodiment.

[0126] (Fourth embodiment) The immunochromatographic test kit 103 according to the fourth embodiment basically has the same configuration and produces the same effects as the immunochromatographic test kit 100 according to the first embodiment, but differs from the immunochromatographic test kit 100 in that it is configured as a linked body in which multiple immunochromatographic test kits 100A to 100C are linked in a direction perpendicular to the development direction D, as shown in Fig. 18. The detection targets of the immunochromatographic test kits 100A to 100C are different from one another.

[0127] The immunochromatographic test kits 100A to 100C may be fixed to each other by any method, for example, by adhesion.

[0128] According to the immunochromatographic test kit 103 of the fourth embodiment, three types of target substances can be detected by immunochromatography using one immunochromatographic test kit 103 configured as a conjugate.

[0129] Example 1 In this example, the results of observing the detection unit 3 of the immunochromatographic test kit 100 using an electron microscope will be described.

[0130] <Sample> The carrier 6 was a porous material made of nitrocellulose. Each detection part 3 was formed to include the bottom surface 7A of the recess 7 using an application needle 21 having a truncated cone-shaped tip with a tip diameter of 50 μm.

[0131] <Observation method> FIG. 19 is a microscopic image obtained by observing the detection section 3 of the sample using a microscope.

[0132] As shown in Figure 19, in the microscope image, a difference occurs between the background and the detection area 3 due to the focus shift caused by the recess 7, so the bottom surface 7A of the recess 7 can be easily located, and the tiny detection area 3 including the bottom surface 7A can be easily located.

[0133] Example 2 In this example, the results of immunochromatography performed using the immunochromatography test kit 100 will be described.

[0134] <Sample> The specimen dropping portion 1 and the conjugate portion 2 were glass fiber pads. The carrier 6 was a porous material made of nitrocellulose. Each detection portion 3 was formed to include the bottom surface 7A of the recess 7 using an applicator needle 21 with a truncated cone-shaped tip and a tip diameter of 50 μm.

[0135] <Testing method> The test method was the same as the test method for the immunochromatographic test kit described above.

[0136] FIG. 20 is an electron microscope image obtained by observing the detection portion 3 of the sample using an electron microscope.

[0137] As shown in Figure 20, the electron microscope image shows a contrast of light and dark due to the recess 7, making it easy to locate the bottom surface 7A of the recess 7 and the tiny detection area 3 including the bottom surface 7A.

[0138] Moreover, FIG. 21 is an electron microscope image of the detection unit 3 before the auxiliary liquid is dropped, and FIG. 22 is an electron microscope image of the detection unit 3 after the auxiliary liquid is dropped.

[0139] It was confirmed that the image shown in FIG. 22 has clearer edges than the image shown in FIG.

[0140] The embodiments and examples disclosed herein are illustrative in all respects and should not be considered limiting. The scope of the present invention is defined by the claims rather than the above description, and it is intended to include meanings equivalent to the claims and all modifications within the scope thereof. [Explanation of symbols]

[0141] 1 sample dropping section, 2 conjugate section, 3 detection section, 3A first detection section, 3B second detection section, 3C third detection section, 4 control section, 5 absorption section, 6 carrier, 7 recess, 7A bottom surface, 7B wall surface, 11 X-axis table, 12 Y-axis table, 13 Z-axis table, 14 coating mechanism, 15 observation optical system, 16 camera, 17 operation panel, 18 monitor, 19 control computer, 20 coating needle holder, 21 coating needle, 22 coating material container, 41 servo motor, 42 stand, 43 cam, 44 bearing, 45 cam connecting plate, 46 moving part, 47 coating needle holder fixing section, 48 coating needle holder storage section, 49 linear guide, 50 spring, 51, 52 fixing pin, 70 Coating materials, 100, 100A, 100C, 101, 102, 103 Immunochromatography test kits.

Claims

1. A method for manufacturing an immunochromatographic test kit comprising a sample application section onto which a sample is dropped, a conjugate section to which a labeled antibody having the property of binding to a target substance in the sample is attached, and a detection section to which a capture antibody having the property of binding to the target substance is attached, The process includes forming the detection unit on the carrier, A method for producing an immunochromatographic test kit, comprising the step of forming the detection unit, which includes forming a concave shape on the carrier having a bottom surface and wall surfaces facing each other on either side of the bottom surface, and simultaneously coating the concave shape with the capture antibody.

2. The method for manufacturing an immunochromatographic test kit according to Claim 1, wherein in the step of forming the detection unit, the concave shape having a tapered shape such that the distance between the opposing wall surfaces narrows as it approaches the bottom surface is formed.

3. The method for manufacturing an immunochromatographic test kit according to claim 1 or 2, wherein in the step of forming the detection unit, the detection unit is formed which is not observable by the naked eye but is observable by a microscope.

4. The method for producing an immunochromatographic test kit according to claim 3, wherein the width of the bottom surface is equivalent to the field of view of an electron microscope capable of directly observing a labeled substance with a size of 1 μm or less that is bound to the labeled antibody.

Images