Inspection tool and inspection kit

Abstract

To provide an inspection tool that does not require an expensive and complicated device for inspection and is capable of more easily confirm detection than conventional chromatography-based inspection kits, and an inspection kit including this inspection tool.SOLUTION: Provided is an inspection tool comprising: a chromatography body having a sample pad for absorbing a sample, a conjugate pad on which a labeled antibody is loaded, a membrane, and an absorption pad for preventing backflow of the sample; and a chromatography container for accommodating the chromatography body. The chromatography container has an observation opening on the surface side of the chromatography body, and has a daylight opening at a position corresponding to the observation opening, so that the light emitted by the labeled antibody can be confirmed from the observation opening by applying light from the daylight opening.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a test tool and a test kit including the same for detecting various targets, such as biomarkers derived from living organisms, such as antibodies, antigens, proteins, and nucleic acids, and extracellular substances, such as chemical molecules. More specifically, the present invention relates to a test tool and a test kit including the same that do not require complicated devices for detection and that can perform detection and confirmation more easily than conventional chromatography-based test tools. [Background technology]

[0002] Many test kits have been proposed for detecting various targets such as antibodies, antigens, proteins, biomarkers, chemical molecules, nucleic acids, etc. Among them, test tools using immunochromatography are widely used due to their ease of use, allowing judgment by visual inspection, and various types have been proposed. For example, Patent Document 1 proposes a solvent-containing substance testing kit that can eliminate the scattering of test substances due to spillage of the test liquid from the test container during testing, can prevent floating substances from entering the test container during testing, and can easily check the test results and improve the accuracy of the determination. Specifically, since the opening of the squeezeless bottle is sealed by a sealing lid during testing, even if the bottle is accidentally knocked over, the sample will not spill and the target substance will not scatter, contaminating the surrounding environment, and floating substances will not enter the bottle from the opening and adversely affect the test results. In addition, since the detection zone is located above the bottle, a cover required in conventional products is not required, and a test kit is proposed that improves the visibility and accuracy of the determination results. Patent Document 2 also proposes an inspection device and cartridge capable of reducing the risk of erroneously identifying the color state of the inspection area. Specifically, the proposed inspection device includes a loading section in which the cartridge is loaded, an illumination section having an emission end for emitting illumination light that illuminates the inspection area whose color state changes depending on whether the specimen contains a test substance, and a detection section for optically detecting the color state of the inspection area irradiated with the illumination light, the cartridge includes a strip with an inspection area provided therein, and a case in which the strip is housed and an observation window for observing the inspection area from the outside is formed on the surface, the emission end is disposed at a position separated from the observation window on the back surface of the case, the detection section is disposed at a position facing the observation window on the front surface side of the case, and the illumination light penetrates from the back surface of the case into the inside of the case and reaches the inspection area while being diffusely reflected on the inner surface of the case, thereby illuminating the inspection area from the front surface side. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent Publication No. 2021-156809 [Patent Document 2] JP 2023-32867 A Summary of the Invention [Problem to be solved by the invention]

[0004] However, with the testing tools proposed above, the chromatography results cannot be clearly confirmed visually, and therefore a detection device must be used to read them, which poses the problem that detection and confirmation cannot be easily performed, as it requires expensive and complicated equipment.

[0005] Therefore, an object of the present invention is to provide a test tool and a test kit including the same that do not require expensive and complicated equipment for detection and enable detection and confirmation more easily than conventional chromatography-based test kits. [Means for solving the problem]

[0006] As a result of intensive research to solve the above problems, the inventors discovered that the above object can be achieved by applying detection light directly from the back side of the observation surface to the position on the membrane where the labeled antibody is to be detected. As a result of further research based on this discovery, they also discovered that the above object can be achieved, and have thus completed the present invention. That is, the present invention provides the following inventions. 1. A test tool comprising a chromatographic body having a sample pad for absorbing a sample, a conjugate pad carrying a labeled antibody, a membrane, and an absorption pad for preventing backflow of the sample, and a chromatographic container for storing the chromatographic body, The chromatography vessel has an observation opening on the surface side of the chromatography vessel body, and a light collecting opening at a position corresponding to the observation opening, and is configured so that light emitted by the labeled antibody can be confirmed from the observation opening by applying light from the light collecting opening. An inspection tool characterized by the above. 2. The testing tool according to 1, wherein the membrane is laminated on a backing sheet, and the total thickness of the membrane and the backing sheet is 160 to 510 μm. 3. The inspection tool according to 1, which has a filter on its front side that transmits light in a predetermined wavelength range. The test tool described in 4.1, A light source that irradiates light in the visible light region (300 nm to 770 nm), The wavelength region of the light emitted by the light source is 300 to 400 nm. Test kit. 5. The testing tool according to 1, wherein the labeled antibody contains a fluorescent substance. 6. The inspection tool according to claim 1, wherein the light collecting opening is provided in a portion corresponding to the line for detection, and the size of the light collecting opening is formed with a width 1 to 3 mm larger than the width of the line. Effect of the Invention

[0007] The testing tool of the present invention does not require complicated devices for detection, and allows detection and confirmation to be performed more easily than conventional chromatography-based testing tools. The test kit of the present invention includes the test tool of the present invention, and therefore can perform detection and confirmation in a shorter time than conventional test kits. [Brief description of the drawings]

[0008] [Figure 1] FIG. 1 is an exploded perspective view showing a schematic overall configuration of a first embodiment of a testing tool of the present invention. [Diagram 2] FIG. 2 is a perspective view showing the testing tool shown in FIG. 1 in an assembled state. [Diagram 3] FIG. 3 is a schematic diagram showing a mode of use of the testing tool shown in FIG. [Figure 4] FIG. 4 is a schematic diagram (photograph substituting a drawing) showing the results of Examples 1 and 2. [Diagram 5] FIG. 5 is an exploded perspective view (corresponding to FIG. 1) that shows a schematic overall configuration of a testing tool according to a second embodiment of the present invention. [Explanation of symbols]

[0009] 1 Test kit, 10 chromatograph body, 11 Sample pad, 13 Conjugate pad, 15 Membrane, 16 Backing sheet, 17 Absorption pad, 20 chromatograph container, 21 Upper half, 23 Observation opening, 25 Sample insertion port, 27 Lower half, 29 Light collection opening, 30 Light source DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] The present invention will now be described in further detail. As shown in Figures 1 and 2, a testing tool 1 according to one embodiment of the present invention comprises a sample pad 11 for absorbing a sample, a conjugate pad 13 carrying a labeled antibody, a chromatography body 10 having an absorption pad 17 and a membrane 15 for sucking up the electrophoresed sample and preventing backflow, and a chromatography container 20 for housing the chromatography body 10. Further details are provided below.

[0011] [Detection target] The detection targets in the detection kit of this embodiment include antibodies, antigens, proteins, nucleic acids, and other biomarkers that can be test targets, as well as other compounds. [Chromatography unit] The chromatographic body 10 includes a membrane 15, which is a member for separating and visualizing components, which are the main functions of chromatography, a conjugate pad 13 with a labeled antibody 40 (see FIG. 3) attached thereto, an absorbent pad 17 for absorbing excess liquid, and a sample pad 11 placed on the end of the conjugate pad 13. The membrane 15 is also provided with a test line 15a and a control line 15b in a conventional manner. The shapes and thicknesses of these members can be formed in the same manner as members used in normal immunochromatography. In this embodiment, all of them are rectangular, and the thicknesses can be any thickness that can be normally adopted without any particular restrictions. The materials for forming these members can be any materials normally used for this type of chromatography without any particular restrictions, and in this embodiment, the membrane 15 is laminated on a backing sheet 16 and is formed from materials such as nitrocellulose and additives such as a surfactant, and the conjugate pad 13, the absorbent pad 17, and the sample pad 11 are formed from nonwoven fabrics made of cellulose fibers. The conjugate pad may be formed from glass fibers. It is preferable that the membrane 15 is of a material and has a thickness that allows it to emit light well when illuminated with light from a light source and that allows it to be clearly visually observed through an observation opening, which will be described later. From this viewpoint, examples of the fiber material forming the membrane include, but are not limited to, cellulose, nitrocellulose, polyolefin, polyethylene, polyethylene terephthalate, cellulose acetate, polyvinylidene difluoride (PVDF), glass fiber, nylons, etc. As the additive, any surfactant that is usually mixed or coated in this type of chromatographic membrane can be used without any particular limitation. The fiber material is important from the viewpoint of obtaining the desired effect of the present invention, and it is particularly preferable to use nitrocellulose as the fiber material. The backing sheet may be any sheet that has been conventionally used in this type of chromatography without any particular limitation, and its thickness is the same as that of the conventional sheet. The thickness of the membrane is usually expressed as the total thickness including the backing sheet, and in the present invention, the total thickness is preferably 160 to 510 μm. The total thickness usually has a range, for example, as shown in the examples, such as 180 (minimum thickness) to 220 μm (maximum thickness) (Example 2) or 215 to 255 μm (Example 1). Taking this into consideration, the minimum thickness is preferably 160 to 250 μm, more preferably 200 to 230 μm. The maximum thickness is preferably 200 to 510 μm, more preferably 240 to 300 μm. From the viewpoint of visibility from the front side (arrow direction in FIG. 2), it is preferable to set it within the above range. In particular, it is important to find that the minimum thickness is preferably 160 μm or more in the present invention. Although a thinner thickness would seem to show better visibility from the viewpoint of light transmission, the inventors' investigations have revealed that a certain degree of thickness is preferable. The reason for this is unclear, but it is believed that a certain degree of thickness provides a good balance between the flow of liquid and the retention of the reaction product with the labeling drug, thereby improving visibility. The membrane preferably has a water flow rate of 50 to 300 seconds / 4 cm, more preferably 55 to 280 seconds / 4 cm. The water flow rate is measured as follows. A 5 cm long membrane is cut out and a mark is placed at the 4 cm position. One end of the membrane is immersed in ultrapure water (0.1 mL), and the time (seconds) it takes for the ultrapure water to reach the 4 cm position is measured. In addition, the chromatographic body of this embodiment may be provided with a case or cover for covering the above-mentioned components, although this is not shown in the figures. Also, components that are advantageous for visual observation, such as a fluorescent filter and a bandpass filter, may be provided. In particular, it is preferable to use a filter that transmits only light of a specific wavelength depending on the labeled antibody used. For example, when the labeled antibody is a europium-based fluorescent substance, a filter that transmits mainly (at least 90% of the light amount) light in the wavelength range of 615±10 nm can be used. In this embodiment, two types of lines are shown, but the present invention is not limited to this, and three or more types of lines may be provided. Moreover, instead of the rectangular shape as in this embodiment, various shapes such as a circular shape may be adopted.

[0012] [Chromatography vessel] The chromatography vessel 20 has an observation opening 23 on the front side of the chromatography body 10, and a light collecting opening 29 at a position facing the observation opening 23 on the back side, and is configured so that light emission of the labeled antibody can be confirmed from the observation opening 23 by shining light through the light collecting opening 29. Specifically, as shown in Figs. 1 and 2, the chromatography vessel 20 of this embodiment 8 is composed of a plate-shaped upper half 21 and a box-shaped lower half 27 with an opening on the top. The upper half 21 has an observation opening 23 at a position corresponding to the test line 15a and the control line 15b, and a sample insertion port 25 at a position corresponding to the sample pad 11. The lower half 27 has a light collecting opening 29 at a position corresponding to the observation opening 23. The size of the light collecting opening 29 is not particularly limited as long as it is located and has a size that allows light to be supplied to the test line 15a and the control line 15b, but from the viewpoint of maintaining the accuracy of detection, it is preferable to make it the minimum size including the test line 15a and the control line 15b. Here, "corresponding position" refers to a position where the light passing through the light-collecting opening directly hits test line 15a and control line 15b, and a position where the observation opening functions to allow the light from the labeled antibody to be clearly observed at test line 15a and control line 15b without any obstruction. The material for forming the chromatography container is not particularly limited, and various materials such as metals and plastics can be used, but plastics are preferably used because of their good moldability and light weight.

[0013] [Labeled antibody] In this embodiment, the labeled antibody carried on the conjugate pad 13 is not particularly limited, but it is preferable that the labeled antibody emits light with sufficient brightness for detection when irradiated with light from the light-collecting opening on the back side. As such a labeled antibody, a mixture of a detection antibody and a fluorescent substance is preferably used. As the fluorescent substance, in particular, the fluorescence brightness (brightness at the wavelength of the emitted fluorescence) is preferably 0.8 or more when the brightness of the excitation light (light at the maximum excitation wavelength) is taken as 1, and more preferably in the range of 0.9 to 1.5 in order to visually confirm the light from the fluorescent substance. Note that the fluorescent substance is not limited, and any labeled antibody that can be visually confirmed from the front side (the direction of the arrow in Figure 2) can be used without any particular limitation. Specific examples of the fluorescent substance include rare earth fluorescent complexes, and examples of the rare earth complexes include europium complexes, terbium complexes, samarium complexes, and dysprosium complexes. In particular, one or more compounds selected from the group consisting of europium complexes, terbium complexes, samarium complexes, and dysprosium complexes can be preferably used as the fluorescent substance. Among these, europium complexes are preferred in terms of obtaining the desired effects of the present invention, and specific examples of europium complexes include the following compounds. Sodium [4'-(4'-amino-4-biphenylyl)-2,2':6',2"-terpyridine-6,6"-diylbis(methyliminodiacetate)]europate(III) (ATBTA-Eu 3+ ),N 1 -(p-isothiocyanatobenzyl)-ethylenediaminetetraacetic acid Eu 3+ Complex (SCN-Ph-EDTA-Eu 3+ ), N 1 -(p-isothiocyanatobenzyl)-diethylenetriamine-N 1 ,N 2 ,N 3 ,N 3 -Eu tetraacetate 3+ Complex (SCN-Ph-DTTA-Eu 3+ ) Eu(β-NTA)3(TOPO)2(10 -14 (detection limit at M level) A so-called fluorescence enhancement solution containing β-NTA-TOPO-Triton X-100 must be added. Eu of 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid 3+ Complex (BCPDA-Eu 3+ ) is used as a labeling agent. Chlorosulfonylated tetradentate β-diketone compound 4,4'-bis(1”,1”,1”,2”,2”,3”,3”-heptafluoro-4”,6”-hexanedione-6”-yl)chlorosulfo-o-terphenyl (BHHCT) BHHCT-Eu 3+ Complex EDTA-Tb 3+ Ternary complexes of with salicylic acid derivatives Two types of fluorescent labeling agents: tris(bipyridine cryptate)-Eu 3+ (Fluorescence energy transfer donor dye, TBP-Eu 3+ ) and allophycocyanin (fluorescence energy transfer acceptor dye, cross-linked allophycocyanin, a pigment protein with a molecular weight of 104 kD, a maximum fluorescence emission wavelength of 665 nm, and a fluorescence quantum yield of approximately 0.7, abbreviated as XL665). BCPDA-Eu 3+ Labeled polyvinylamine-biotin streptavidin complex Combination of poly(Glu:Lys)-streptavidin conjugate and europium fluorescent complex-labeled poly(Glu:Lys)-BSA-streptavidin conjugate 4-[2-(4-isothiocyanatophenyl)ethynyl]-2,6-bis{[N,N- bis(carboxymethyl)amino]methyl}pyridine} and Eu 3+ Those in a state where they are encapsulated in complex particles with europium, for example, polystyrene latex nanoparticles containing europium fluorescent complexes etc. can be mentioned. The following commercially available products can also be used. Estapor (registered trademark) europium microsphere series (Merck) Europium Conjugation Kit (ab269889, Abcam) FS Eu beads (manufactured by Tamagawa Seiki Co., Ltd.), etc. The amount of the labeled antibody used is arbitrary depending on the object to be detected, but it is preferable to adjust the amount of the fluorescent substance to be in the range of 1 to 20, particularly preferably 8 to 12, per one object to be detected. This adjustment is determined according to the expected amount of the object to be detected (calculated from the content of the object to be detected predicted relative to the amount of the collected sample). In addition, it is also possible to use two or more types of fluorescent substances in combination. In addition, the above-mentioned complexes can be used as they are as the fluorescent substance. Furthermore, it is preferable to use the above-mentioned complexes in a state in which they are encapsulated in particles, from the viewpoints of dispersibility in the membrane and visual visibility of the fluorescence. For example, it is preferable to use the above-mentioned commercially available product Estapor (registered trademark) europium microsphere series (manufactured by Merck, see Examples). The mixing ratio of the detection antibody to the fluorescent substance is not particularly limited, but when particles (polymer particles, etc.) encapsulating the complex are used, the detection antibody / particle (mg / g) is preferably 20 to 80, more preferably 40 to 80, and most preferably 40 to 60. Various types of labeled antibodies can be used depending on the thickness and material of the membrane, and are not limited to the specific examples given above.

[0014] <Detection kit> The detection kit of this embodiment comprises the detection tool of the present invention of the above embodiment and a light source. 〔light source〕 The light source used in this embodiment can be one that can irradiate light in the visible light region (300 to 770 nm), but it is particularly preferable to use one that can irradiate light with a wavelength of 300 to 400 nm. There are no particular limitations on the light source that can irradiate light with such a wavelength, and commercially available products can be used. in particular, Alonefire, product name "SV16 Small 5W Ultraviolet Blacklight Wavelength 365nm USB Rechargeable UV LED Light"; Portable small black light keychain for glowing original puzzles (UV-LED375-nano) manufactured by Nichia Chemical, product name "1-lamp LED black light" etc. can be used without particular limitation. Depending on the labeled antibody used, a light source in the visible light range can also be used.

[0015] [Other components] In addition to the above-mentioned chromatography body and chromatography vessel, the detection tool and detection kit of the present invention can be used in combination with various other members without departing from the spirit of the present invention. Furthermore, when used as one tool in a detection kit, other tools can be used in addition to the light sources described above without departing from the spirit of the present invention. [Color level sample] The other tools may include, for example, a color level sample. The color level sample is for enabling the detection kit of the present embodiment to perform not only qualitative analysis but also quantitative analysis. For example, chromatography is performed according to a plurality of concentrations of the labeled detection target, light is applied from a light source to each concentration to emit light, the fluorescent membranes are aligned, color samples corresponding to the plurality of concentrations are created, and these are compiled. That is, the fluorescent color level sample includes fluorescent color samples corresponding to the concentrations of the plurality of detection targets. The number and degree of the plurality of concentrations to be aligned is arbitrary depending on the detection target, but it is usually preferable to have about 10 to 20 types, which is useful for quantitative judgment. By providing a fluorescent color level sample in this manner, a detection kit capable of quantification can be constructed by visually checking the color level of the fluorescent color level sample and the fluorescent color of the chromatography.

[0016] [Manufacturing method] The detection tool of this embodiment is obtained by manufacturing a chromatographic body according to a conventional method, manufacturing an upper half and a lower half by a conventional processing method, storing the chromatographic body in the obtained lower half, and finally fitting the upper half into the upper opening of the obtained half. The chromatographic body of this embodiment can be obtained as follows. The detection antibody is applied to the membrane in a line shape and dried to form the test line 15a and the control line 15b. As the above antibody, any antibody that can be detected by this type of chromatography, such as SARS-CoV-2 Nucleoprotein antibody, anti-HA antibody, and anti-mouse antibody, can be used as the detection target without any particular restrictions. In addition, detection substances other than the above antibodies can also be used. Thereafter, the antibody bound to the labeled antibody is applied to a conjugate pad and dried to obtain a conjugate pad, and the chromatographic body can be formed. Next, the sample pad, conjugate pad, membrane and absorbent pad are formed in the usual manner and superimposed in the manner shown in FIG. Furthermore, a plurality of such chromatographic bodies are prepared, and sample solutions adjusted to a plurality of concentrations are dropped onto each chromatographic body to perform immunochromatography. After a predetermined time (10 to 30 minutes) has elapsed, light is irradiated from a light source, and the light from the observation opening at that time is photographed to photograph a plurality of emitted lights, and a color level sample can be created by assembling the photographs of the plurality of lights.

[0017] [Usage and effects] The detection tool and detection kit of this embodiment can be used as shown in Fig. 3. In Fig. 3, a labeled antibody 40 is shown typically, and a sample and an object to be detected are also shown typically for explanation. 2, to use the detection kit 1 of this embodiment, first, a sample 41 is dropped onto the sample pad 11. The sample 41 moves from the conjugate pad 13 to the membrane 15 by capillary action. At this time, the detection target 43 in the sample 41 reacts with the labeled antibody 40 supported on the conjugate pad 13 and moves to the membrane 15 in a bound state. The bound product of the labeled antibody 40 and the detection target 43 reacts with the antibody constituting the test line 15a and remains at the test line 15a. Then, light of a predetermined wavelength is emitted from light source 30 through light collection opening 29 on the back side, and detection can be performed by visually observing through observation opening 23 on the front side. At this time, it is also possible to measure the concentration of the detection target 43 by preparing a color level sample and comparing the color of the color level sample with the color of the light observed visually through the observation opening 23. When this detection procedure is performed in a dark place, such as in a dark box (using the detection kit of the present invention), the desired effects of the present invention can be obtained more effectively.

[0018] Since the detection tool and detection kit of this embodiment are configured as described above, it is possible to detect the presence of the antibody by irradiating the light-collecting opening with light from the light source and visually inspecting the observation opening. In particular, the light-collecting opening allows the light from the light source to be applied directly to the membrane from a close distance, making it possible to sufficiently judge the presence of the antibody by visual inspection. In particular, when the fluorescent substance described above is used as the labeled antibody, it is possible to confirm the fluorescence of a different color tone from the light from the light source, making it easier to judge the presence of the antibody by visual inspection. In addition, since light is applied from the back, it is possible to use an optimal filter to obtain the wavelength required for visual inspection without having to consider the influence of light irradiated from the viewing side, making visual judgment more accurate and easier. In addition, because light is applied from the back side and the fluorescence is observed from the front side, a light source with a wide wavelength range can be used. Conventionally, not only the desired fluorescence but also light of other wavelengths such as autofluorescence was emitted as noise, and since the viewing side and the irradiated side were the same, it was difficult to efficiently remove the noise. However, in the present invention, it is possible to efficiently remove such noise and only view the light of the desired wavelength. By using two types of filters, one on the incident light side and one on the reflected light side, the excitation light and the reflected light can be efficiently selected, and the wavelengths required for visual observation can be more easily recognized and selected. In this sense, the membrane can also be sandwiched between filters. In this embodiment, a filter is installed in the light collection opening 29 on the back side, and light is irradiated from the back side by the light source, but at that time, only light of the wavelengths required for fluorescence can be irradiated, and fluorescence can be efficiently produced. On the other hand, by installing a filter in the observation opening 23 on the front side, light that can be considered noise can be removed, resulting in good visual observation. Visual confirmation is preferable because it allows for easy testing, but conventional test kits that allow for easy visual judgment have not been very accurate. The reason is that visual judgment is difficult due to background. Conventionally, in order to eliminate the background, two types of filters, one for excitation light and one for incident light, were required, which made the device configuration of the excitation light incident device on the visual inspection cassette complicated, and it was difficult to operate by installing two types of filters and shining light on them to read the results. As a result of extensive research into how to solve this problem, it was discovered that the above-mentioned problem could be solved by applying light to the back of the membrane, and extensive research was conducted into constructing a chromatography vessel suitable for this case, and furthermore, a more preferable detection antibody (fluorescent substance) was investigated, leading to the completion of the present invention. That is, by configuring as in this embodiment and the second embodiment, the desired effect of the present invention can be achieved by "providing an opening" and "transmitting light", and judgment can be performed simply by placing it over a light source, and since it is easy to see, accurate judgment can be performed.

[0019] Next, a second embodiment of the present invention will be described with reference to Fig. 5. In the description of the second embodiment, differences from the embodiment shown in Fig. 1 described above will be described in detail. Therefore, the reference numerals in the embodiment shown in Fig. 1 are numbered with 100, and descriptions of structural parts such as members configured in the same manner as in the embodiment shown in Fig. 1 and the positional relationship of each member will be omitted. Therefore, points that are not particularly described are the same as the embodiment shown in Fig. 1. 5, the observation openings 123a, 123b and the light collecting openings 129a, 129b in the chromatography container 120 are different. That is, the observation opening 123a and the light collecting opening 129b are provided at a position corresponding to the test line 115a, and the observation opening 123b and the light collecting opening 129b are provided at a position corresponding to the control line 115b. The observation openings 123a, 123b and the light collecting openings 129a, 129b are each 1 to 3 mm larger (wider) than the width of the corresponding test line 115a and the control line 115b (meaning that when the width of each line is w, the width w' of each opening is w+1 to 3 mm).

[0020] As in the second embodiment, by providing only an opening of the minimum size (width) required according to the width of each line, the influence of the background can be minimized, and more accurate qualitative and quantitative judgment can be made by visual inspection. In addition, the operation for judgment is also simplified because it is easier to read by visual inspection. Note that the test kit of this embodiment is preferably manufactured by making the entire kit and then providing each opening, which avoids the problem of error between the lines and the openings.

[0021] The present invention is not limited to the above-described embodiment, and various modifications are possible without departing from the spirit of the present invention. EXAMPLES

[0022] [Examples 1 and 2] Estapor (registered trademark) microspheres (trade name "K1-050") were used for visual confirmation to verify the feasibility of visual use. Separately, FS Eu COOH beads (trade name "TAB5849N2140") were used as a labeling substance to obtain the detection kit shown in Figure 1 according to the above-mentioned manufacturing method. The raw materials and light sources used are shown below. Sample pad: Product name "CF4" (manufactured by Cytiva) Conjugate pad: Product name "33glass" (manufactured by Cytiva) Membrane: Example 1: Product name "FF80 HP Plus Thick" (manufactured by Cytiva), total thickness including backing: 215 to 255 μm, water flow rate: 60 to 95 seconds / 4 cm Example 2: Product name "FF80 HP" (manufactured by Cytiva), total thickness including backing (substituted with top cover tape, product name "BDPET25(A)PL Thin 8LK") is 180 to 220 μm, water flow rate is 60 to 95 seconds / 4 cm Absorbent pad: Product name "SureWick (registered trademark)" cellulose sample pad (C083 type, manufactured by Merck) Microsphere concentration: antibody:particle 1:10, particle equivalent 0.006%, 10μL Light source: Alonefire product name "SV16 small 5W ultraviolet black light wavelength 365nm" Using the obtained detection kit (using FS Eu COOH beads (product name "TAB5849N2140") as a labeling substance), H5 subtype avian influenza antigen (hemagglutinin) was detected. For detection, five antigen solutions were prepared with concentrations of 0, 10, 100, 1000, 10000, and 100000 (unit: pg / mL), and 0.1 mL was dropped onto each test kit (the actual amount used is shown in the figure) and the test was performed. As a result, it was possible to visually judge the color by shining light from a light source through the light collection opening. Furthermore, since the luminescence level differs depending on the antigen concentration in the antigen solution, it is possible to perform simple, but quantitative, detection by comparing with a level sample. A comparison of the sensitivities is shown in Figure 4. As is clear from the results shown in Figure 4, both Examples 1 and 2 had high sensitivity, but assuming visual inspection, it can be seen that Example 1 is better able to perform visual inspection than Example 2. Furthermore, a detection kit shown in Fig. 5 was obtained using the configuration of Example 1 (Example 3). The obtained detection kit was tested in the same manner as in Example 1, and visually judged. As a result, compared to Example 1, there was less background influence, the color of the fluorescence could be recognized, and a better judgment could be made.

Claims

1. A test tool comprising a chromatography body having a sample pad for absorbing a sample, a conjugate pad carrying a labeled antibody, a membrane, and an absorption pad for preventing backflow of the sample, and a chromatography container for storing the chromatography body, The chromatography vessel has an observation opening on the surface side of the chromatography vessel body, and a light collecting opening at a position corresponding to the observation opening, and is configured so that light emitted by the labeled antibody can be confirmed from the observation opening by applying light from the light collecting opening. An inspection tool characterized by the above.

2. 2. The testing tool according to claim 1, wherein the membrane is laminated on a backing sheet, and the total thickness of the membrane and the backing sheet is 160 to 510 μm.

3. A filter that transmits light in a specified wavelength range is provided on the front side. The testing tool according to claim 1.

4. The testing tool according to claim 1 ; A light source that irradiates light in the visible light region (300 nm to 770 nm), The wavelength region of the light from the light source is 300 to 400 nm. Test kit.

5. 2. The test tool according to claim 1, wherein said labeled antibody contains a fluorescent substance.

6. 2. The inspection tool according to claim 1, wherein the light collecting opening is provided in a portion corresponding to a line for detection, and the size of the light collecting opening is formed with a width 1 to 3 mm larger than the width of the line.

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