JP7712622B2 - Manufacturing method of immunochromatography test kit - Google Patents

Description

The present invention relates to a method for producing an immunochromatographic test kit.

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.

In conventional immunochromatography, the presence or absence of color development in the detection section of the immunochromatography test kit (also called the development support or chromatographic medium) is visually observed. Specifically, when the detection target bound to the labeled antibody is captured by the capture antibody fixed to the detection section and sufficiently accumulates on the detection section, the color development due to the labeled antibody is visually confirmed.

International Publication No. 2010/061772 (Patent Document 1) discloses an immunochromatographic test kit in which the detection unit is formed with a size of several millimeters or more that can be visually determined.

International Publication No. 2010/061772

Methods for forming the detection section of the immunochromatographic test kit described above include the dispenser method and the inkjet method. In order to form a detection section with a small size of less than a few millimeters using these methods, it is necessary to make the opening width of the nozzle tip smaller.

However, if the liquid containing the capture antibody is highly viscous, clogging can occur if the opening width at the nozzle tip is reduced.

For this reason, depending on the viscosity of the liquid, it is difficult to reduce the size of the detection unit, and it is not possible to reduce the amount of expensive capture antibody used, making it difficult to reduce costs.

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.

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

In the manufacturing method of the immunochromatographic test kit, the outer diameter of the tip of at least one application needle may be less than 1 mm.

In the manufacturing method of the immunochromatographic test kit, in the step of forming and applying at least one detection portion, the capture antibody may be applied in a dot shape using at least one application needle.

In the manufacturing method of the 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.

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

In the manufacturing method of the immunochromatographic test kit, the multiple coating needles may include a first coating needle having at its tip a first capture antibody having a property of binding to a first type of detection target, and a second coating needle having at its tip a second capture antibody having a property of binding to a second type of detection target. In the step of forming the multiple detection sections, the first coating needle may be used to coat the first region with the first capture antibody to form the first detection section, and the second coating needle may be used to coat the second region with the second capture antibody to form the second detection section.

In the manufacturing method of the immunochromatographic test kit, the multiple coating needles may include a first coating needle having at its tip a first capture antibody having a property of binding to a first type of detection target, a second coating needle having at its tip a second capture antibody having a property of binding to a second type of detection target, and a third coating needle having at its tip a third capture antibody having a property of binding to a third type of detection target. In the step of forming the multiple detection parts, the first coating needle may be used to coat the first region with the first capture antibody to form the first detection part, the second coating needle may be used to coat the second region with the second capture antibody to form the second detection part, and the third coating needle may be used to coat the third region with the third capture antibody to form the third detection part.

In the manufacturing method of the immunochromatographic test kit described above, the sample dropping section, the conjugate section, and at least one detection section may be formed on a single porous member.

In the manufacturing method of the immunochromatographic test kit, the sample dropping section, the conjugate section, and at least one detection section may be formed on multiple porous members.

The present invention provides 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.

1 is a perspective view of an immunochromatographic test kit according to a first embodiment. FIG. 2 is a partially enlarged plan view of a detection section of the immunochromatographic test kit shown in FIG. 1 . FIG. 4 is a partial enlarged view for explaining the width of a detection portion. FIG. 4 is a partially enlarged cross-sectional view taken along the line IV-IV in FIG. 2. 1 is a flowchart of a manufacturing method of an immunochromatographic test kit according to the first embodiment. 1 is a perspective view of a coating device used in a manufacturing method for an immunochromatographic test kit according to embodiment 1. FIG. FIG. 7 is a front view of a coating unit of the coating apparatus shown in FIG. 6 . FIG. 7 is a side view of the application unit of the application apparatus shown in FIG. 6 . 7 is a front view illustrating the operation of a coating unit of the coating apparatus shown in FIG. 6. 7 is a front view illustrating the operation of a coating unit of the coating apparatus shown in FIG. 6. 3 is a partially enlarged plan view showing a modified example of the detection unit shown in FIG. 2. FIG. 3 is a partially enlarged plan view showing another modified example of the detection unit shown in FIG. 2. FIG. 13 is a partially enlarged plan view of a detection section of the immunochromatographic test kit according to embodiment 2. FIG. 14 is a partially enlarged plan view showing a modified example of the detection unit shown in FIG. 13. FIG. 14 is a partially enlarged plan view showing another modified example of the detection unit shown in FIG. 13. FIG. 14 is a partial enlarged plan view showing still another modified example of the detection unit shown in FIG. 13. FIG. FIG. 11 is a perspective view of an immunochromatographic test kit according to a third embodiment. FIG. 13 is a perspective view of an immunochromatographic test kit according to a fourth embodiment. 4 is an electron microscope image obtained by observing the detection unit 3 of Example 1 using an electron microscope. 13 is an electron microscope image obtained by observing the detection unit 3 of Example 2 using an electron microscope. 13 is an electron microscope image obtained by observing the detection unit 3 with an electron microscope before the auxiliary liquid is dropped in Example 2. 13 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.

Below, an embodiment of the present invention will be described as an example with reference to the drawings. Note that in the following drawings, the same or corresponding parts are given the same reference numbers, and their description will not be repeated.

(Embodiment 1)
<Immunochromatography test kit>
1 and 2, the immunochromatographic test kit 100 according to the first embodiment mainly includes 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 Fig. 1 and Fig. 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.

The specimen dripping section 1 is the portion onto which the specimen is dripped. The specimen dripping section 1 is formed of a porous material such as a glass fiber pad, a cellulose fiber pad, or a polyester pad.

The conjugate portion 2 is a portion to which a labeled antibody having a property of binding to a detection target in a sample is fixed. The labeled antibody is a conjugate of an antibody that specifically recognizes and binds to a first portion (first epitope) of the detection target and a labeling substance. The antibody can be arbitrarily selected according to the detection target. The labeling substance can be arbitrarily selected from labeling substances used in conventional immunochromatography. The labeling substance includes 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-developing fine particles such as liposomes containing a coloring agent. The labeling substance includes at least one selected from the group consisting of precious metal nanoparticles such as gold nanoparticles, platinum nanoparticles, platinum-gold nanoparticles, and silver nanoparticles, titanium nanoparticles, iron nanoparticles, nickel nanoparticles, and cadmium nanoparticles, as metal nanoparticles. The metal nanoparticles may be colloidal metal nanoparticles having a particle size of 1 nm or more and 100 nm or less. 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 the same.

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

As shown in FIG. 2, the out-of-plane shape of each detection unit 3 is dot-shaped. 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 with 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 with a size of 1 μm or less that bind to labeled antibodies. The dot width of each detection unit 3 is, for example, 100 μm or less.

The width of the detection section 3 is measured as follows. First, a sample containing a sufficient amount of the detection target is dropped onto the sample dropping section 1 to induce an antigen-antibody reaction on the immunochromatographic test kit 100. Next, an electron microscope image of the detection section 3 is obtained according to the test method described below. When the detection section 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 obtained by focusing on the bottom surface 7A is used. Next, the outer shape of the area in which the labeled antibody is observed in the image is specified as the outer shape of the detection section 3. Next, the width of the outer shape of the detection section 3 is measured. As shown in FIG. 3, when the outer shape of the area in which the labeled antibody is observed undulates inward and outward, the diameter D1 of the circumscribing circle CC of the outer shape and the diameter D2 of the inscribing circle IC of the outer shape are calculated by image processing, and the intermediate value between the diameters D1 and D2 is defined as the width of the outer shape of the detection section 3.

The detection units 3 shown in FIG. 2 are spaced apart from one another 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.

The planar shape of each detection unit 3 is not particularly limited as long as it is dot-shaped, but is, for example, circular. The planar shape of each detection unit 3 may also be elliptical, square, rectangular, or the like.

As shown in FIG. 4, a plurality of recesses 7 are formed in the detection region t of the carrier 6. Each recess 7 is recessed with respect to the upper surface of the carrier 6. Each recess 7 has a bottom surface 7A and a wall surface 7B connecting the bottom surface 7A and the upper surface of the carrier 6, and the opening of the upper 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 becomes narrower as it approaches the bottom surface 7A, but the opening diameter of the upper surface 6A and the diameter of the bottom surface 7A may be equal. The width of the bottom surface 7A of each recess 7 is the minimum size that can be confirmed by visual observation, 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 directly observes a labeled substance of 1 μm or less that binds to a labeled antibody. For example, the dot width of each bottom surface 7A is 100 μm or less. The depth of each bottom surface 7A is deeper than the focal depth of an electron microscope. The depth of each bottom surface 7A is, for example, 10 μm or more.

The planar shape of each bottom surface 7A is, for example, circular. The planar shape of each bottom surface 7A may also be elliptical, square, rectangular, or the like.

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, for example, simultaneously in the carrier 6. A method for simultaneously forming the detection unit 3 and the recess 7 in the carrier 6 will be described in detail in the manufacturing method of the immunochromatography test kit described below.

The control region c is disposed on the opposite side of the detection region t from the conjugate section 2 in the development direction D. The control region C includes a control section 4. The control section 4 is a section to which a control antibody that binds to the labeled antibody is fixed. The out-of-plane shape of the control section 4 may be any shape, for example, a line shape.

The control unit 4 is formed on the carrier 6. The control unit 4 is disposed on the opposite side of the detection unit 3 from the conjugate unit 2 in the deployment direction D.

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

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 detection area t with respect to the control area c in the development direction D.

The out-of-plane shape of the carrier 6 is rectangular. The carrier 6 has a longitudinal direction along 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, and may include, 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 a porous member 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, for example, to the porous member constituting the conjugate section 2. Each of the porous members constituting the conjugate section 2 and the absorption section 5 is bonded, for example, to each of the porous members constituting the carrier 6.

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 onto a substrate (not shown) (also called a backing sheet) using an adhesive or an adhesive sheet. The carrier 6 may be a thin film formed on the substrate.

<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.

First, a specimen is dropped onto the specimen dropping section 1. The dropped specimen flows along the developing direction D due to capillary action. The object to be detected in the specimen binds to the labeled antibody in the conjugate section 2. The conjugate of the object to be detected and the labeled antibody moves along the developing direction D together with the labeled antibody that is not bound to the object to be detected due to capillary action. The object to be detected that has bound to the labeled antibody binds to the capture antibody in the detection section 3. As a result, the labeled antibody that has bound to the object to be detected is accumulated in the detection section 3. The labeled substance that has bound to the labeled antibody in the detection section 3 is directly observed using an electron microscope. Furthermore, the labeled antibody that has not bound to the object to be detected binds to the control antibody in the control section 4. As a result, the labeled antibody that has not bound to the object to be detected is captured in the control section 4. The labeled substance that has bound to the labeled antibody in the control section 4 is directly observed using an electron microscope. The excess specimen that has flowed downstream of the control section 4 in the developing direction D is absorbed by the absorption section 5.

A test method using an 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 elapsed, an auxiliary liquid is dropped.

The auxiliary liquid has electrical conductivity to prevent static electricity and heat generation, which contribute to image clarity, and/or the property of polymerizing to form a film under measurement conditions using an electron microscope. 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, as optional components, at least one compound selected from monosaccharides, disaccharides, salts, and buffer solutions.

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, in the microscopic image (bright field image) of the electron microscope, defocus caused by the distance between the top surface of the carrier 6 and the bottom surface 7A is observed. Therefore, by checking the presence or absence of defocus, the recess 7 can be searched for in the microscopic image, and the detection unit 3 can be easily searched for in the image. Then, the focus of the image can be adjusted to the bottom surface 7A, so that the focus of the image can be adjusted to the detection unit 3. Next, an electron microscope image is taken, and the number of labeled substances in the image is measured. Since the contours of the labeled substances can be clearly identified in the electron microscope image, the number of labeled substances in the image can be measured. If the number of measured labeled substances exceeds a predetermined judgment criterion, it is judged that the detection target is present in the sample (i.e., "positive").

The above observations and measurements are performed in the same manner, for example, for multiple 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 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."

<Manufacturing method of immunochromatography test kit>
Next, a description will be given of an example of a manufacturing method of the immunochromatographic test kit 100. As shown in Fig. 5, the manufacturing method of 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.

In the process of preparing the carrier 6, a carrier 6 having a control unit 4 formed on its upper surface, or a carrier 6 having no control unit 4 formed on its upper surface, is prepared.

In the process of forming the multiple detection sections 3, a coating needle 21 (see FIG. 10) holding the capture antibody at its tip is used to coat the top 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 a recess 7 in the carrier 6, and at the same time, the capture antibody is coated on the bottom surface 7A of the recess 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 top 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.

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

<Configuration of Coating Apparatus>
Next, an example of a coating device used for forming the multiple detection units 3 in the manufacturing method of the immunochromatographic test kit 100 will be described.

As shown in FIG. 6, the coating device according to embodiment 1 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.

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 section that is 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 that straddles the Y-axis table 12 in the X-axis direction is fixed to the bottom of the processing chamber.

The X-axis table 11 is placed on a structure that is installed so as to straddle the Y-axis table 12 in the X-axis direction. A moving 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 moving 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 above-mentioned Y-axis table 12 is movable in the Y-axis direction relative to the X-axis table 11.

The Z-axis table 13 is installed on the moving body connected to the X-axis table 11 as described above. 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 be moved in the Z-axis direction.

The operation panel 17, monitor 18, and control computer 19 are used to control the Y-axis table 12, the X-axis table 11, the Z-axis table 13, the observation optical system 15, and the 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.

<Configuration of Coating Mechanism 14>
Next, a detailed configuration of the coating mechanism 14 shown in Fig. 6 will be described. As shown in Fig. 7 and Fig. 8, the coating mechanism 14 mainly includes a servo motor 41, a stand 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 direction along the Z-axis direction shown in Fig. 6. The servo motor 41 is held by the stand 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.

The cam 43 includes a center portion connected to the rotating shaft of the servo motor 41 and a flange portion connected to one end of the center portion. The upper surface of the flange portion (the surface on the servo motor 41 side) is a cam surface. This cam surface is formed in an annular shape along the outer periphery of the center portion, and is formed in a slope 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 the longest distance from the bottom surface of the flange portion, a lower end flat region that is the shortest distance from the bottom surface of the flange portion, and a slope region that connects the upper end flat region and the lower end flat region. The upper end flat region is disposed at a distance from the lower end flat region in the circumferential direction relative to the rotating shaft.

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 against the cam surface of the cam 43 by a spring 50, which will be described later. Therefore, when the cam 43 rotates, the bearing 44 is held in a state in which the outer peripheral surface of the outer ring is 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.

A fixed pin 51 is installed on the stand 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 the spring 50 acts on the bearing 44 via the movable part 46 and the cam connecting plate 45. The tensile force of the spring 50 keeps the bearing 44 pressed against the cam surface of the cam 43.

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.

The coating needle holder 20 is stored in the 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 shape. 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.

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

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

<Operation of Coating Device>
Next, the operation of the coating device in the process of forming the multiple detection units 3 in the manufacturing method of the immunochromatographic test kit will be described. First, the carrier 6 is mounted on 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, and the carrier 6 is placed 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 in which the detection unit 3 is 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 upper surface of the carrier 6 when the coating needle is placed in the second state.

Next, the servo motor 41 of the coating mechanism 14 is operated to switch from the first state to the second state. Specifically, the servo motor 41 is operated to rotate the rotation shaft of the servo motor 41 to rotate the cam 43. As a result, the height of the cam surface of the cam 43 in the Z-axis direction changes, so that the position of the bearing 44 in contact with the cam surface in the Z-axis direction also changes according 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, by operating the servo motor 41, the coating needle 21 moves in the Z-axis direction, and the first state shown in FIG. 9 and the second state shown in FIG. 10 are switched. In the coating mechanism 14, the rotational motion of the servo motor 41 can be converted into motion (up and down motion) 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.

In the first state shown in FIG. 9, the coating needle 21 is positioned at the uppermost position in its range of motion. At this time, the tip of the coating needle 21 is immersed in the coating material 70 held in the coating material container 22. In the first state, the outer ring of the bearing 44 is in contact with the upper flat area of the cam surface of the cam 43.

In the second state shown in FIG. 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 end face and side of the tip of the coating needle 21 hold a coating material containing a capture antibody. 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 top surface of the carrier 6.

In this way, by operating the servo motor 41 to switch from the first state to the second state, a recess 7 is formed in the carrier 6 and the capture antibody is applied to the bottom surface 7A of the recess 7. Next, the servo motor 41 is operated to switch from the second state to the first state.

Furthermore, by operating only the X-axis table 11 and the Y-axis table 12, the area in which the second detection unit 3 is to be formed is positioned directly under the coating mechanism 14, and then the Z-axis table 13 is lowered by a distance d, and 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.

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 effect of the manufacturing method of the immunochromatographic test kit 100 relating to embodiment 1 will be explained in comparison with a manufacturing method in which each detection part 3 is formed using a dispenser (hereinafter simply referred to as Comparative Example 1) and a manufacturing method in which each detection part 3 is formed by an inkjet method (hereinafter simply referred to as Comparative Example 2).

In Comparative Examples 1 and 2, in order to form the detection part 3 in a very small 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 discharged, and clogging may occur. In Comparative Example 2, in a method in which a piezoelectric element or the like is used to instantaneously increase the pressure inside the container to discharge the liquid material containing the capture antibody, the high discharge pressure means that the liquid material tends to spread when it hits the carrier 6, resulting in problems such as an unstable shape of the detection part 3.

In contrast, in the manufacturing method of the immunochromatographic test kit 100, the tip diameter of the coating needle 21 can be made small to form a minute detection unit 3, so clogging does not occur as in Comparative Examples 1 and 2, regardless of the viscosity. Furthermore, in the above manufacturing method, the position of the tip of the coating 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 the variation in size of the detection unit 3 can be reduced, and the immunochromatographic test kit 100 can be stably manufactured.

In addition, in Comparative Example 1, it is difficult to stably form detection parts 3 with dot widths of less than 1 mm in order to apply droplets, and in particular, it is difficult to stably form detection parts 3 with dot widths of less than 100 μm.

In contrast, the manufacturing method of the immunochromatographic test kit 100 uses a coating needle 21, so that the immunochromatographic test kit 100 in which the planar shape of each detection part 3 is dot-shaped and the width of each detection part 3 is less than 1 mm can be easily and stably manufactured.

Furthermore, in the manufacturing method of the immunochromatographic test kit 100, the recesses 7 are formed in the carrier 6 using the coating 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 section 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 associated with the formation of the detection section 3, and no increase in manufacturing costs.

Next, the effect of the immunochromatographic test kit 100 according to the first embodiment will be described 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 originating from the labeled antibody can be visually confirmed. Therefore, in Comparative Example 1, a sufficient area of the detection part (e.g., several ^mm2 or more) is required to prevent erroneous judgment by visual judgment, and it is difficult to reduce the amount of expensive capture antibody fixed to the detection part. As a result, it is difficult to reduce the manufacturing cost in Comparative Example 1.

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 application 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, so the amount of capture antibody fixed to each detection unit 3 can be reduced compared to the amount of capture antibody fixed to 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.

Furthermore, with 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 counted, allowing the substance to be detected in the sample to be quantitatively evaluated.

Furthermore, when the detection unit formed on a flat surface is miniaturized as in Comparative Example 1, it is necessary to enlarge the detection unit with a microscope or the like to confirm the labeled antibody, but it is difficult to search for and align the tiny detection unit with a microscope. One possible solution to this problem is to provide a marker on the kit and then provide the detection unit at a specified distance from that marker. However, in this case, equipment for providing the marker is required in addition to the equipment for forming the detection unit, and the number of processes increases, making it impossible to reduce the manufacturing takt time and making it difficult to reduce costs.

In contrast, in the immunochromatographic test kit 100, multiple 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.

Furthermore, by focusing the image on the bottom surface 7A, the image can be easily focused on the detection unit 3. In other words, 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.

In addition, unlike Comparative Example 1, in the immunochromatographic test kit 100, there is no need to provide a separate marker to be used when searching for the detection unit 3, and there is no need to prepare equipment for providing the marker separately from the 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 the case where a marker is provided in Comparative Example 1.

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 fixed to each detection part 3 is significantly reduced compared to the amount of capture antibody fixed to each detection part in Comparative Example 3, so that the manufacturing cost of the immunochromatographic test kit 100 can be significantly reduced compared to the manufacturing cost of Comparative Example 3.

The immunochromatographic test kit 100 includes multiple detection units 3. Compared to an immunochromatographic test kit that includes only one detection unit 3, this immunochromatographic test kit 100 allows the labeled substances of the multiple detection units 3 to be confirmed in an electron microscope image, improving the reliability of the test results.

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

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

<Modification 1>
The immunochromatographic test kit 100 according to the first embodiment can be modified as follows.

The detection area t may include at least one detection unit 3 .
In the immunochromatographic test kit 100, the detection units 3 have the same configuration, but are not limited to this. In the immunochromatographic test kit 100, the concentration of the capture antibody fixed to each detection unit 3 is constant, but the concentration of the capture antibody fixed to each detection unit 3 may be different from each other. For example, the concentration of the capture antibody may be gradually decreased from one side to the other side in a direction perpendicular to the development direction D. In addition, a set of patterns consisting of a plurality of detection units 3 having different concentrations of the capture antibody may be arranged side by side. Such a plurality of detection units 3 can be easily formed by using a plurality of application mechanisms 14 having different concentrations of the capture antibody in the application material stored in the application material container 22.

In addition, at least one of the planar shape of each detection unit 3, the dot width of each detection unit 3, the distance between two adjacent detection units 3, the planar 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 coating needles 21 or multiple types of coating conditions.

In the immunochromatographic test kit 100, multiple recesses 7 are formed in the detection region t of the carrier 6, and one detection section 3 includes the bottom surface 7A of one recess 7, but this is not limited to this.

Each detection unit 3 may include the entire bottom surface 7A of each recess 7 and at least a portion of the wall surface 7B. Each detection unit 3 may also include the entire bottom surface 7A and wall surface 7B of each recess 7. Each detection unit 3 may also 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.

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 manufacturing method by moving the coating material 70 attached to the tip or tip of the coating needle 21 upward without being pushed downward after contacting the upper surface of the carrier 6. In addition, 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.

As shown in FIG. 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.

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

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

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, so that the contact time between the tip of the coating needle 21 or the coating material 70 attached to the tip and the carrier 6 can be increased. As a result, the amount of capture antibody applied to the carrier 6 can be increased and the concentration can be increased, so that the antigen-antibody reaction with the detection target can be more stabilized.

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.

(Embodiment 2)
The immunochromatographic test kit 101 of embodiment 2 has basically the same configuration and produces the same effects as the immunochromatographic test kit 100 of 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.

A first capture antibody having a property of binding to a first type of detection target is fixed to the first detection section 3A. A second capture antibody having a property of binding to a second type of detection target different from the first type of detection target is fixed to the second detection section 3B.

As shown in FIG. 13, the multiple detection units 3 include, for example, multiple first detection units 3A and multiple second detection units 3B. The multiple first detection units 3A are arranged at intervals from one another in a direction perpendicular to the deployment direction D. The multiple first detection units 3A are arranged, for example, one-dimensionally. The multiple second detection units 3B are arranged at intervals from one another in a direction perpendicular to the deployment direction D. The multiple second detection units 3B are arranged, for example, one-dimensionally.

The multiple first detection units 3A and multiple second detection units 3B are spaced apart 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.

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.

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.

Each first detection portion 3A includes the bottom surface 7A of each recess 7. Each second detection portion 3B includes the bottom surface 7A of each recess 7.

The immunochromatographic test kit 101 according to the second embodiment can detect two types of detection targets from a single sample by immunochromatography.

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 section 3A by coating a first capture antibody on the carrier 6, and a second coating mechanism 14 for forming a second detection section 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.

In the manufacturing method of the immunochromatographic test kit 101, the first coating needle 21 holds a first capture antibody at its tip. The second coating needle 21 holds a second capture antibody at its tip. In the process of forming the multiple detection sections 3, the first detection section 3A is formed by applying the first capture antibody to a first region of the carrier 6 using the first coating needle 21. Furthermore, the second detection section 3B is formed by applying the second capture antibody to a second region of the carrier 6 using the second coating needle 21.

By manufacturing the immunochromatographic test kit 101 using such an applicator, the number of manufacturing steps can be reduced compared to when the immunochromatographic test kit 101 is manufactured using a single applicator equipped with only one applicator mechanism 14, and the manufacturing costs can be reduced compared to when the immunochromatographic test kit 101 is manufactured using multiple applicators equipped with only one applicator mechanism 14.

<Modification 2>
The immunochromatographic test kit 101 according to the second embodiment may also have the same modifications as those of the immunochromatographic test kit 100 according to the first embodiment.

Furthermore, the immunochromatographic test kit 101 may have the following modifications.
The plurality of first detection units 3A and the plurality of second detection units 3B may be arranged alternately.

As shown in Figures 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. Each third detection unit 3C has a third capture antibody immobilized thereon that has the property of binding to a third type of detection target that is different from the first type of detection target and the second type of detection target.

As shown in FIG. 14, the multiple first detection units 3A, the multiple second detection units 3B, and the multiple third detection units 3C are arranged alternately, for example, in a direction perpendicular to the deployment direction D.

As shown in FIG. 15, the multiple first detection units 3A, the multiple second detection units 3B, and the multiple third detection units 3C are arranged, for example, in a row of multiple units alternating 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, for example, the distance between the adjacent first detection units 3A and second detection units 3B. The distance between two adjacent second detection units 3B and the distance between two adjacent third detection units 3C are shorter than, for example, the distance between the adjacent second detection units 3B and third detection units 3C.

As shown in FIG. 16, the first detection units 3A, the second detection units 3B, and the third detection units 3C are arranged one-dimensionally, for example, in the deployment direction D. Furthermore, the first detection units 3A, the second detection units 3B, and the 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 the distance between 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 the distance between two adjacent third detection units 3C in the deployment direction D.

In the modified examples shown in Figures 14 to 16, in a set of the first detection unit 3A, the second detection unit 3B, and the third detection unit 3C, at least any of the planar shape of each detection unit 3, the dot width of each detection unit 3, the distance between two adjacent detection units 3, the planar 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.

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.

The second detection unit 3B may have a rectangular planar shape, and the first detection unit 3A and the third detection unit 3C may have a circular planar shape.

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

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.

Such a first detection unit 3A, second detection unit 3B, and third detection unit 3C can be easily formed by using multiple types of coating needles 21 or multiple types of coating conditions.

In this way, the first detection unit 3A, the second detection unit 3B, and the third detection unit 3C can be easily identified within one field of view.

(Embodiment 3)
The immunochromatographic test kit 102 according to the third 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 the specimen dropping portion 1 and the conjugate portion 2 are formed on the carrier 6. That is, in the immunochromatographic test kit 102, the specimen dropping portion 1, the conjugate portion 2, the multiple detection portions 3, and the control portion 4 are formed on the upper surface of a single porous member (one carrier 6).

In the immunochromatographic test kit 102 shown in FIG. 17, the absorption section 5 is also formed in the carrier 6.

The specimen dropping section 1 is an area on the carrier 6 where the specimen is dropped, and is located on one side of 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 developing direction D relative to the control area C.

The immunochromatographic test kit 102 has a reduced number of parts compared to the immunochromatographic test kit 100, so the manufacturing costs of the immunochromatographic test kit 102 can be reduced compared to the manufacturing costs of the immunochromatographic test kit 100.

<Modification 3>
The immunochromatographic test kit 102 according to the third embodiment may also adopt the same modified examples as the immunochromatographic test kit 100 according to the first embodiment and the immunochromatographic test kit 101 according to the second embodiment.

(Embodiment 4)
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 the immunochromatographic test kit 103 is configured as a linked body in which a plurality of 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.

Each of the immunochromatographic test kits 100A to 100C may be fixed to each other by any method, for example by adhesive.

According to the immunochromatographic test kit 103 according to the fourth embodiment, three types of detection targets can be detected by immunochromatography using one immunochromatographic test kit 103 configured as a conjugate.

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

<Sample>
The carrier 6 was a porous member made of nitrocellulose. Each detection portion 3 was formed to include the bottom surface 7A of the recess 7 using a coating needle 21 having a truncated cone shape and a tip diameter of 50 μm.

Observation method
FIG. 19 is a microscopic image obtained by observing the detection portion 3 of the above sample using a microscope.

As shown in FIG. 19, in the microscope image, the focus shift caused by the recess 7 creates a difference between the background and the detection area 3, so the bottom surface 7A of the recess 7 can be easily searched, and the tiny detection area 3 including the bottom surface 7A can be easily searched.

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

<Sample>
The specimen dropping portion 1 and the conjugate portion 2 were glass fiber pads. The carrier 6 was a porous member made of nitrocellulose. Each detection portion 3 was formed to include the bottom surface 7A of the recess 7 using a coating needle 21 having a truncated cone shape and a tip diameter of 50 μm.

<Testing method>
The test method was the same as that of the immunochromatographic test kit described above.

Figure 20 shows an electron microscope image obtained by observing the detection portion 3 of the above sample using an electron microscope.

As shown in FIG. 20, the electron microscope image shows a contrast of light and dark due to the recess 7, so the bottom surface 7A of the recess 7 can be easily detected, and the minute detection portion 3 including the bottom surface 7A can be easily detected.

Figure 21 is an electron microscope image of the detection unit 3 before the auxiliary liquid is dripped, and Figure 22 is an electron microscope image of the detection unit 3 after the auxiliary liquid is dripped.

The image shown in Figure 22 shows clearer image edges than the image shown in Figure 21.

The embodiments and examples disclosed herein are illustrative in all respects and should not be considered limiting. The scope of the present invention is indicated by the claims rather than the above description, and is intended to include the meaning equivalent to the claims and all modifications within the scope.

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 Movable section, 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 kit.

Claims (10)

A method for producing an immunochromatographic test kit comprising, on a porous member, a specimen dropping section onto which a specimen is dropped, a conjugate section to which a labeled antibody having a property of binding to a substance to be detected in the specimen is attached, and at least one detection section to which a capture antibody having a property of binding to the substance to be detected is attached,
and applying the capture antibody to the porous member using at least one application needle having the capture antibody at a tip thereof to form the at least one detection portion.
A method for manufacturing an immunochromatographic test kit, in which, in the process of forming at least one detection portion, the at least one application needle is brought into contact with the upper surface of the porous member and then further pushed into the porous member to form a recess in the porous member, while at the same time applying the capture antibody to the bottom surface of the recess . The method further includes the step of providing a coating device;
The coating device includes a coating mechanism,
the coating mechanism includes the at least one coating needle and a servo motor having a rotation shaft;
The method for producing an immunochromatographic test kit according to claim 1 , wherein in the step of forming the at least one detection unit, a depth of the bottom surface of the recess varies depending on an amount of rotation of the rotation shaft of the servo motor. The method for producing an immunochromatographic test kit according to claim 1 or 2, wherein the depth of the recess is 10 μm or more. The method for producing an immunochromatographic test kit according to any one of claims 1 to 3 , wherein the outer diameter of the tip of the at least one application needle is less than 1 mm. The method for manufacturing an immunochromatographic test kit according to any one of claims 1 to 4 , wherein in the step of forming and applying the at least one detection unit, the capture antibody is applied in a dot shape by the at least one application needle. The method for producing an immunochromatographic test kit according to any one of claims 1 to 5, wherein in the step of forming the at least one detection unit, a plurality of detection units are formed by applying the capture antibody one-dimensionally or two -dimensionally at intervals using a plurality of application needles. the plurality of coating needles include a first coating needle carrying at its tip a first capture antibody having a property of binding to a first type of detection target, and a second coating needle carrying at its tip a second capture antibody having a property of binding to a second type of detection target,
7. The method for manufacturing an immunochromatographic test kit as described in claim 6, wherein in the step of forming the multiple detection sections, the first capture antibody is applied to a first region using the first application needle to form a first detection section, and the second capture antibody is applied to a second region using the second application needle to form a second detection section. the plurality of coating needles include a first coating needle holding at its tip a first capture antibody having a property of binding to a first type of detection target, a second coating needle holding at its tip a second capture antibody having a property of binding to a second type of detection target, and a third coating needle holding at its tip a third capture antibody having a property of binding to a third type of detection target,
7. The method for manufacturing an immunochromatographic test kit as described in claim 6, wherein in the step of forming the multiple detection sections, the first capture antibody is applied to a first region using the first application needle to form a first detection section, the second capture antibody is applied to a second region using the second application needle to form a second detection section, and the third capture antibody is applied to a third region using the third application needle to form a third detection section. The method for producing an immunochromatographic test kit according to any one of claims 1 to 8 , wherein the specimen dropping portion, the conjugate portion, and the at least one detection portion are formed on a single porous member. The method for producing an immunochromatographic test kit according to any one of claims 1 to 8 , wherein the specimen dropping portion, the conjugate portion, and the at least one detection portion are formed on a plurality of the porous members.