Antigen detection kit and detection method

Abstract

To provide a kit that enables more reliable and quicker detection of allergens, and a detection method.SOLUTION: A kit for detecting antigens is provided, having a protein layer containing a protein which may be casein, bean-derived protein, oat-derived protein, or rice koji-derived protein. The protein layer also contains a water retention agent, protease activator, and antimicrobial agent.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to kits and methods for detecting antigens. [Background technology]

[0002] In recent years, allergy-related diseases have been increasing and become a social problem. In order to prevent allergies, it is important to detect allergens present in the environment. As a method for detecting allergens, a petri dish having a gel containing a polypolypeptide placed on a base layer is placed in a humid box for a predetermined period of time, stained, and the allergen attached to the gel is detected. The gel is decomposed by proteases in the allergens attached to the gel, and the decomposed part of the base layer is exposed, thereby detecting the allergens. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 2012-21980 A Summary of the Invention [Problem to be solved by the invention]

[0004] In the above-mentioned methods, it may take a long time to detect allergens.

[0005] An object of the present disclosure is to provide a kit and a detection method capable of detecting allergens more reliably and in a shorter period of time. [Means for solving the problem]

[0006] The present disclosure includes the following aspects. [Section 1] 1. A kit for detecting an antigen, the kit having a protein layer comprising a protein, the protein being casein, a bean-derived protein, an oat-derived protein, or a rice koji-derived protein. [Section 2] Item 1. The kit according to item 1, wherein the protein layer further comprises a moisture retaining agent. [Section 3] Item 3. The kit according to item 1 or 2, wherein the moisture retaining agent is agar, sodium alginate, polyacrylic acid, polyvinyl alcohol, polyacrylamide, carrageenan, pectin, or glycerin. [Section 4] Item 4. The kit according to item 3, wherein the moisture retaining agent is a combination of glycerin and at least one selected from agar, sodium alginate, polyacrylic acid, polyvinyl alcohol, polyacrylamide, carrageenan, and pectin. [Section 5] Item 5. The kit according to any one of items 1 to 4, wherein the protein layer is white, milky brown, or brown. [Section 6] Item 6. The kit according to any one of Items 1 to 5, wherein the protein layer contains a protease activator. [Section 7] Item 7. The kit according to any one of Items 1 to 6, wherein the protein layer contains a cysteine ​​protease inhibitor, a serine protease inhibitor, an aspartic acid protease inhibitor, or a metalloprotease inhibitor. [Section 8] Item 8. The kit according to any one of Items 1 to 7, wherein the protein layer comprises an antimicrobial agent. [Section 9] Item 9. The kit according to any one of items 1 to 8, wherein the thickness of the protein layer is 1 μm to 3 mm. [Section 10] Item 10. The kit according to any one of Items 1 to 9, wherein the antigen is a cysteine ​​protease antigen, a serine protease antigen, an aspartic acid protease antigen, or a metalloprotease antigen. [Section 11] Item 11. The kit according to any one of items 1 to 10, comprising a support member and a monolayer of the protein layer. [Section 12] Item 12. The kit according to item 11, wherein the support member is a petri dish. [Section 13] Contacting an antigen with the kit according to any one of items 1 to 12; Detecting digestion marks on the surface of the kit; A method for detecting an antigen, comprising: [Section 14] Item 14. The method of claim 13, wherein the antigen comprises a cysteine ​​protease antigen, a serine protease antigen, an aspartic protease antigen, or a metalloprotease antigen. [Section 15] Item 15. The method according to item 13 or 14, comprising placing the kit according to any one of items 1 to 12 at a detection site and then adding a reaction stopper to the kit. [Section 16] Item 16. The method of item 15, wherein the reaction quenching agent is an alcohol. Effect of the Invention

[0007] According to the kit and method of the present disclosure, it is possible to provide a kit and a detection method that can detect allergens more reliably and in a shorter period of time. [Brief description of the drawings]

[0008] [Figure 1] FIG. 1 shows the observation results 10 minutes after the start of the test using Kit A in the Example. [Diagram 2] FIG. 2 shows the results of observation 30 minutes after the start of the test using Kit A in the example. [Diagram 3] FIG. 3 shows the observation results 60 minutes after the start of the test using Kit A in the Example. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] Hereinafter, the embodiments of the present invention will be described in detail. However, the embodiments shown below are merely examples of kits and detection methods for embodying the technical ideas of the present invention, and the present invention is not limited to the kits and detection methods shown below. The upper and lower limits of the numerical ranges described in this specification can be arbitrarily selected and combined.

[0010] (Antigen detection kit) The present disclosure provides a kit for detecting an antigen having a protein layer that includes a protein.

[0011] The kit of the present disclosure is placed at a location where the presence of an antigen (e.g., an allergen) is to be tested, and the antigen can be detected by observing the digestion marks on the protein-containing layer, specifically, the color change from colored (typically white) to transparent. Specifically, in the kit of the present disclosure, when the protein-containing layer is contacted with a protease derived from the antigen, the protein in the protein-containing layer is hydrolyzed by the protease activity, and the colloidal particles of the protein are decomposed to form transparent sites.

[0012] (antigen) Antigens that can be detected by the kits of the present disclosure are antigens that contain proteases (eg, protease-containing allergens).

[0013] The protease antigen contained in the antigen is not particularly limited, but examples thereof include cysteine ​​proteases, serine proteases, aspartic acid proteases, metalloproteases, trypsin-like proteases, chymotrypsin-like proteases, papain-like proteases, elastase-like proteases, peptidase-like proteases, collagenase-like proteases, and the like.

[0014] Protease-containing antigens can be, for example, insects, insect feces, and insect carcasses; and fragments thereof, where insects can be, for example, mites, cockroaches, etc.

[0015] Insects, insect droppings, and insect carcasses; as well as their debris, contain Der f1 / p1, Der f3 / p3, Der f6 / p6, or Der f9 / p9 in mites and Bla g2 in cockroaches.

[0016] The protease-containing antigen may be, for example, a fungus (mold) or a bacterium, etc. Here, the bacterium may be, for example, Bacillus subtilis, Staphylococcus aureus, etc.

[0017] Fungi (molds) and bacteria contain proteases such as Proteinase K in the case of Tritirachium albulim and subtilisin in the case of Bacillus subtilis.

[0018] The protease-containing allergen may be, for example, pollen, etc. Here, the plants from which the pollen is derived may be, for example, cedar, cypress, ragweed, pine, grass, mugwort, white birch (Betula platyphylla), alder, Alnus alpina, and Obayashabushi.

[0019] Pollen contains many proteins that are highly homologous to pollen allergens of other plants, such as Act c1 homologous protein and Cuc m1 homologous protein in the case of cedar pollen.

[0020] The protease-containing allergen may be, for example, house dust, etc. House dust is a mixture of various allergens, and may be a mixture of the above-mentioned various allergens (insects, insect feces, insect corpses, and their fragments; fungi (molds) and bacteria, etc.; pollen, etc.) in addition to dander and hair derived from humans and non-human animals.

[0021] Among house dust, dander and hair derived from humans and non-human animals may have serine proteases and cysteine ​​proteases derived from mites, molds, and fungi attached to their surfaces.

[0022] (protein-containing layer) The protein-containing layer is a layer that contains a protein.

[0023] The protein is casein, elastin, bean-derived protein, oat-derived protein, or rice koji-derived protein, preferably casein.

[0024] The pulse-derived protein may be a protein derived from soy milk, or a protein derived from almonds, peas.

[0025] The above-mentioned soy milk-derived protein, oat-derived protein, almond-derived protein, rice koji-derived protein, and pea-derived protein are preferably autoclaved before use.

[0026] The autoclave conditions were as follows: Temperature: 50 to 150°C, preferably 80 to 130°C, for example 100 to 120°C Pressure: 1 to 3 atmospheres, preferably 1.5 to 2.5 atmospheres Time: 5 to 60 minutes, preferably 10 to 30 minutes

[0027] As the casein, casein itself or a material containing casein such as skim milk or cow's milk can be used.

[0028] Since the protein-containing layer contains a polypeptide protein that serves as a protease substrate, the protein is decomposed by the protease activity derived from the allergen, and the protein aggregates disappear due to the decomposition, forming a transparent region. The allergen can be detected by the presence or absence of the formation of such a transparent region.

[0029] The amount of protein contained in the protein-containing layer is not particularly limited, but may be, for example, 0.01 to 10 wt %, preferably 0.1 to 1.0 wt %, and more preferably 0.3 to 0.8 wt %, calculated as the ratio of substrate protein in 100 wt % of the protein-containing layer.

[0030] The protein-containing layer may further contain a moisture-retaining agent, such as agar, sodium alginate, a water-containing polymer, carrageenan, pectin, glycerin, or a water-containing gel (e.g., polyacrylic acid, polyvinyl alcohol, polyacrylamide). The protein-containing layer contains agar. When the protein-containing layer contains a moisture-retaining agent, hydrolysis of the protein is promoted, and allergens can be detected more reliably and in a shorter time.

[0031] The content of the moisture-retaining agent may be, for example, 1 to 100 parts by mass, preferably 10 to 90 parts by mass, more preferably 30 to 70 parts by mass, and even more preferably 40 to 60 parts by mass, relative to 100 parts by mass of protein. Note that this content includes the amount of water.

[0032] In a preferred embodiment, the moisture retaining agent is a combination of glycerin and at least one selected from agar, sodium alginate, polyacrylic acid, polyvinyl alcohol, polyacrylamide, carrageenan, and pectin. By combining glycerin as a moisture retaining agent, it is possible to suppress the loss of moisture in the gel, and the storage stability of the kit is improved.

[0033] In the above embodiment, the content of glycerin in the protein-containing layer may be preferably 1.0% by mass to 30% by mass, more preferably 2.0% by mass to 20% by mass, and even more preferably 2.5% by mass to 10% by mass. By making the content of glycerin 1.0% by mass or more, it is possible to more effectively suppress the loss of moisture. By making the content of glycerin 30% by mass or less, it is possible to suppress the decrease in detectability.

[0034] The protein-containing layer preferably has a certain degree of shape retention. When the protein-containing layer has shape retention, the shape of the transparent region formed in the protein-containing layer is easily retained, so that the protease activity and therefore the amount of antigen can be quantitatively evaluated. In particular, the size of the transparent region allows a relative comparison of the amount of antigen.

[0035] The protein-containing layer may contain a crosslinking agent, which improves the shape retention of the protein-containing layer.

[0036] The crosslinking agent is not particularly limited as long as it can crosslink the gel that is a protein support, and examples thereof include glutaraldehyde, glutaraldehyde oligomer, succinaldehyde, and formaldehyde.

[0037] In one embodiment, the protein-containing layer may contain a protease activator or a protease inhibitor that specifically acts on various proteases contained in the antigen. By containing the protease activator in the protein-containing layer, the hydrolysis reaction of the protease can be promoted, and thus the test time can be shortened. By using the protease activator or the protease inhibitor, it is possible to confirm whether or not a specific protease is present in the sample, and thus it is possible to identify the antigen.

[0038] An example of a protease activator is DTT.

[0039] Protease inhibitors include cysteine ​​protease inhibitors (eg, E-64), serine protease inhibitors (eg, AEBSF), aspartic protease inhibitors, or metalloprotease inhibitors.

[0040] The protein-containing layer may be colored. The protein-containing layer may be colored from the beginning or may be dyed with a dye. The protein layer is preferably white, milky brown, or brown, more preferably white.

[0041] The dyes include red dyes, blue dyes, etc., such as Nylon Red, Nylon Blue, Brilliant Blue, Methylene Blue, Lanasin Black, Biebrich Scarlet, etc. The dye may preferably be Biebrich Scarlet.

[0042] The protein-containing layer may further contain an antimicrobial agent, which can inhibit the growth of microorganisms after use of the kit.

[0043] Examples of the antimicrobial agent include benzalkonium chloride, coconut alkyl dimethyl benzyl ammonium chloride (cation F2-50), Proxel IB, 2-pyridinethiol, 1-oxide, sodium salt (Marucaside SYconc), methyl parahydroxybenzoate (methyl paraoxybenzoate), and the like, with Marucaside SYconc being preferred.

[0044] The amount of the antimicrobial agent added may be preferably 0.01% to 5.0% by mass, more preferably 0.05% to 3.0% by mass, and even more preferably 0.1% to 1.0% by mass, relative to the protein-containing layer.

[0045] In this specification, "colored" means having some color (including white), and colored may be transparent or opaque. For example, it is sufficient if the color is such that the presence of a protein-containing layer can be recognized by visual inspection.

[0046] The thickness of the protein-containing layer may be preferably 1 μm to 3 mm, more preferably 3 μm to 20 μm. The smaller the thickness of the protein-containing layer, the easier it is to form holes in the protein-containing layer, making detection quicker and easier. Also, the smaller the thickness of the protein-containing layer, the darker the color of the protein-containing layer, making it easier to detect the transparent area.

[0047] Unit area of ​​protein-containing layer (1 cm 2 The amount per unit area may be preferably 0.01 to 0.50 g, more preferably 0.03 to 0.30 g, and even more preferably 0.05 to 0.10 g.

[0048] The protein-containing layer may be a single layer or multiple layers, for example two layers. The protein-containing layer is preferably a single layer.

[0049] In one embodiment, the kit of the present disclosure comprises a support member and a monolayer of said protein layer.

[0050] The support member is a member that supports the protein-containing layer and the base layer.

[0051] The support member is particularly limited as long as it is a member capable of supporting the protein-containing layer, and may be a petri dish, a plate, paper, etc. Also, it may be a combination of two or more of a petri dish, a sheet, a plate, paper, etc. For example, it may be a combination of a petri dish and a sheet.

[0052] The support member may contain a fluorescent agent or a luminescent agent. For example, when the support member is a petri dish, a fluorescent agent or a luminescent agent may be applied to the surface of the support member. Alternatively, a transparent petri dish may be combined with a fluorescent or luminescent sheet. When the support member contains a fluorescent agent or a luminescent agent, the base layer is preferably transparent. When the support member contains a fluorescent agent or a luminescent agent, the contrast with the protein-containing layer is increased, making it easier to detect the transparent region.

[0053] The fluorescent agent has a fluorescent effect of absorbing ultraviolet rays from sunlight or artificial light, converting them into visible light, and radiating the visible light. The fluorescent agent is preferably a fluorescent brightening agent. The fluorescent brightening agent can increase the whiteness and blueness of the base layer.

[0054] Examples of fluorescent agents include 7-diethylamino-4-methylcoumarin, 4,4'-bis(2-sulfostyryl)biphenyl disodium, dialkylaminocoumarin, bisdimethylaminostilbene, bismethylaminostilbene, 4-alkoxy-1,8-naphthalenedicarboxylic acid-N-alkylimide, bisbenzoxazolylethylene, 1,4-bis(2-benzoxazolyl)naphthalene, and dialkylstilbene.

[0055] The fluorescent agent may be a commercially available fluorescent agent. Commercially available fluorescent agents include, for example, Super Glo DF-300 (manufactured by Marktec), Kayahor 3BS Liq., Kayahor TAC Liq., Kayaphor HBC Liq., Kayalight B, Kayaphor AS 150, Kayaphor CR 200, Kayaphor JB Liq., Mikawhite ATN conc. (manufactured by Nippon Kayaku Co., Ltd.), TINOPAL OB, TINOPAL NFW. Liqid (manufactured by BASF), Hakkol P, Hakkol PHD, Hakkol RF, Hakkol PSR, Hakkol PSR-B, Hakkol CHP-B, Hakkol PY-2000, Hakkol PYB-D, Hakkol BE conc, Hakkol SG conc 150, Hakkol TH-100, Hakkol S-100, Hakkol RX-1, Hakkol OW-11, Hakkol OW-10extra, Hakkol KIP H / C, Hakkol SF, Hakkol JC, and Hakkol PMS H / C (manufactured by Showa Chemical Industry Co., Ltd.), Nikkafluor SB conc, Nikkafluor KB, Nikkafluor OB, and Nikkabright CX H / C (manufactured by Nippon Chemical Industry Co., Ltd.).

[0056] The fluorescent agents may be used alone or in combination.

[0057] The luminescent agent is a material that absorbs sunlight or artificial light and continues to emit light for a predetermined period of time after the absorption. The luminescent agent can cause the base layer to emit light, thereby enhancing the contrast with the protein-containing layer.

[0058] Examples of the luminescent agent include sulfide-based luminescent agents and aluminate-based luminescent agents. Specific examples of the sulfide-based luminescent agents include CaS:Bi, CaSrS:Bi, and ZnCdS:Cu. Specific examples of the aluminate-based luminescent agents include RMg2Al. 16 O 27 :Eu, RMg2Al16O27 :Eu,Mn,RMg2Al 10 O 17 :Eu, RMg2Al 10 O 17 :Eu,Mn (each R is independently Sr or Ba), SrAl2O4:Eu, SrAl2O4:Eu,Dy, Sr4Al 14 O 25 :Eu, Sr4Al 14 O 25 :Eu,Dy, CaAl2O4:Eu,Nd, etc.

[0059] The luminescent agents may be used alone or in combination.

[0060] (Method of detecting antigen) The present disclosure relates to contacting an antigen with the kit of the disclosure; Detecting traces of digestion on the surface of the kit; Includes.

[0061] (Contacting the kit with the antigen) In the detection methods of the present disclosure, the kit is contacted with the antigen.

[0062] The kit may be contacted with the antigen by placing the kit at the detection site, or the antigen may be collected at the detection site and then contacted. For example, dust may be collected with an air sampler and the collected dust may be inoculated into the kit. The detection site refers to the location where the presence of the antigen is to be confirmed.

[0063] When the kit of the present disclosure has a moisture supply layer, the hydrolysis reaction by the protease can proceed without any special operation, except for placing the kit at the detection site.

[0064] When the kit of the present disclosure has a moisture-permeable layer, for example, hydrolysis reaction by protease can proceed simply by contacting the base layer with water.

[0065] In one embodiment, after contacting the kit with the antigen, the kit may be placed in a water vapor atmosphere. In another embodiment, after contacting the kit with the antigen, the kit may be placed in a location where water is present, for example, on a sponge impregnated with water. By placing the kit in a water vapor atmosphere or in a location where water is present, hydrolysis reaction by protease is promoted.

[0066] (Detecting traces of digestion on the surface of the kit) In the detection method of the present disclosure, a digestion trace is detected on the surface layer of the kit. The surface layer is typically a protein-containing layer. The digestion trace means a transparent area of ​​the protein-containing layer generated by a hydrolysis reaction caused by a protease.

[0067] The detection method of the transparent region is usually performed with the naked eye, but is not particularly limited thereto, and for example, the transparent region may be magnified and observed using a telescope, an overhead projector, a scanner, etc. Alternatively, the transparent region can be detected by acquiring an image of the protein-containing layer and analyzing the image.

[0068] According to the method of the present disclosure, the amount of antigen in a sample can be quantitatively evaluated by summing up the bottom surface area of ​​the transparent regions formed in the protein-containing layer. In addition, the amount of antigen in a sample can be quantitatively evaluated by summing up the number of transparent regions formed in the protein-containing layer. Alternatively, the amount of antigen in a sample can be quantitatively evaluated based on both the sum of the bottom surface area of ​​the transparent regions and the total number of the transparent regions.

[0069] The area of ​​the transparent region may be measured, for example, by binarizing a photographed image of the protein-containing layer using computer processing, and measuring the area of ​​the region corresponding to the transparent region as the number of pixels.

[0070] In a preferred embodiment, the method of the present disclosure may include adding a reaction stopper to the kit after a predetermined time has elapsed since the kit is placed at the detection location. The reaction of the antigen can be stopped by adding a reaction stopper to the kit. If the reaction is not stopped, the digestion trace becomes blurred and difficult to observe after a long time, but stopping the reaction can suppress the blurring of the digestion trace. In addition, the antigen can be quantified by measuring the exact area of ​​the digestion trace at a predetermined time, or by measuring the color (or shade) of the digestion trace. Quantification based on the color of the digestion trace is performed by creating a calibration curve (color value-quantitative value) in advance at a predetermined allergen concentration and fitting the color of the digestion trace to be measured to the calibration curve. It is preferable to convert the color of the digestion trace into a numerical value.

[0071] The reaction stopper may be preferably alcohol or a protease inhibitor, more preferably alcohol. By using alcohol as the reaction stopper, the whiteness of the protein layer is improved and the digestion traces are more easily observed. In addition, by using alcohol or a protease inhibitor as the reaction stopper, the reaction can be stopped without destroying the protein.

[0072] The alcohol may be preferably an alcohol having 1 to 6 carbon atoms, more preferably methanol, ethanol, or isopropanol.

[0073] The alcohol is preferably added as an aqueous solution. The alcohol concentration may be preferably 10% by mass to 80% by mass, more preferably 30% by mass to 70% by mass, and further preferably 40% by mass to 60% by mass.

[0074] The protease inhibitor can be appropriately selected depending on the antigen, and can be, for example, a serine protease inhibitor, a cysteine ​​protease inhibitor, an aspartic acid protease inhibitor, or a metalloprotease inhibitor.

[0075] The amount of the reaction stopper added is preferably an amount capable of covering the surface of the kit.

[0076] The timing for adding the reaction stopper may be when the antigen reaction has progressed and traces of digestion become clear, for example, 10 to 2 hours, preferably 20 minutes to 1 hour, after the kit has been placed at the detection site.

[0077] The kit and detection method of the present disclosure have been described in detail above. Note that the kit and detection method of the present disclosure are not limited to those exemplified above. EXAMPLES

[0078] The present disclosure will be described in further detail below with reference to examples, but the present disclosure is not limited to the following examples.

[0079] Example 1 1.0 g of skim milk powder (190-12865, manufactured by Wako) was added to 50 mL of 1 M Tris-HCl aqueous solution and dispersed by stirring at room temperature. Next, 0.5 g of agar (016-11875, manufactured by Wako) was added and further stirred at 120°C for 2 hours to dissolve. The temperature of the solution was increased to 70°C and further stirred for 30 minutes, dithiothreitol (DTT) was added to a final concentration of 2 mM, and further stirred for 5 minutes. 4 mL of the obtained casein solution was added to a plastic petri dish with a diameter of 90 mm, left to stand and solidified, forming a casein-containing layer, and antigen detection kit A was prepared.

[0080] Example 2 30 mL of milk was stirred at 120°C for 15 minutes, then 0.3 g of agar (WAKO 016-11875) was added and further stirred at 120°C for 30 minutes. The temperature of the solution was increased to 70°C and further stirred for 30 minutes, dithiothreitol (DTT) was added to a final concentration of 2 mM, and further stirred for 5 minutes. 5 mL of the obtained casein solution was added to a 90 mm diameter plastic petri dish, left to stand and solidified, forming a casein-containing layer, to prepare antigen detection kit B.

[0081] Example 3 0.08 g of casein and 120 μL of 4% NaOH aqueous solution were added to 25 mM Tris-HCl (pH 6.8) and stirred at 70°C. 0.12 g of casein and 160 μL of 4% NaOH aqueous solution were added, heated to 120°C and stirred for 30 minutes. 0.1 g of casein was added and stirred. 0.3 g of agar (WAKO 016-11875) was then added and stirred for 60 minutes. Dithiothreitol (DTT) was then added to a final concentration of 2 mM and stirred for another 5 minutes. 5 mL of the obtained casein solution was added to a plastic petri dish with a diameter of 90 mm, left to stand and solidified, forming a casein-containing layer, to prepare antigen detection kit C.

[0082] Comparative Example 1 An agar solution was prepared by mixing 50 mL of 1 M Tris-HCl aqueous solution, 1 g of agar, and 5 g of talc. The obtained agar solution was added to a plastic petri dish having a diameter of 90 mm, and allowed to stand to gel, forming a base layer. A gelatin solution was prepared by mixing 10 mL of 1 M Tris-HCl aqueous solution, 0.5 g of gelatin, 20 μL of 1 M DTT, 10 μL of 25% glutaraldehyde, 3 mL of ethanol, and 0.03 g of Biebrich scarlet. The gelatin solution was added onto the base layer to form a polypeptide-containing layer, and antigen detection kit a was prepared.

[0083] (Experimental Example) Trypsin was dissolved in 25 mM Tris-HCl (pH 6.8) on the antigen detection kit obtained above, and the solution was further diluted to 0.1 mg / mL. 1 μL of the obtained solution was applied to the kit. After 10 minutes, 30 minutes, and 60 minutes from the dropwise addition, the presence or absence of digestion traces on the casein-containing layer was observed. The visibility of the digestion traces was evaluated. The evaluation criteria are as follows. For reference, images of the sample of Kit A after 10 minutes, 30 minutes, and 60 minutes are shown in Figures 1 to 3, respectively. 1: It was difficult to visually observe the digestive traces. 2: Digestion marks were visible. 3: Digestion marks were clearly visible. [Table 1] [Industrial Applicability]

[0084] The antigen detection kit and detection method of the present disclosure are preferably used for detecting antigens, particularly allergens.

Claims

1. 1. A kit for detecting an antigen, the kit having a protein layer comprising a protein, the protein being casein, a bean-derived protein, an oat-derived protein, or a rice koji-derived protein.

2. The kit of claim 1 , wherein the protein layer further comprises a moisture retention agent.

3. 2. The kit of claim 1, wherein the moisture retention agent is agar, sodium alginate, polyacrylic acid, polyvinyl alcohol, polyacrylamide, carrageenan, pectin, or glycerin.

4. The kit of claim 3 , wherein the moisture-retaining agent is a combination of glycerin and at least one selected from agar, sodium alginate, polyacrylic acid, polyvinyl alcohol, polyacrylamide, carrageenan, and pectin.

5. The kit of claim 1 , wherein the protein layer is white, milky brown, or brown.

6. The kit of claim 1 , wherein the protein layer comprises a protease activator.

7. 10. The kit of claim 1, wherein the protein layer comprises a cysteine ​​protease inhibitor, a serine protease inhibitor, an aspartic protease inhibitor, or a metalloprotease inhibitor.

8. The kit of claim 1 , wherein the protein layer comprises an antimicrobial agent.

9. The kit of claim 1, wherein the thickness of the protein layer is from 1 μm to 3 mm.

10. 2. The kit of claim 1, wherein the antigen is a cysteine ​​protease antigen, a serine protease antigen, an aspartic protease antigen, or a metalloprotease antigen.

11. The kit of claim 1 comprising a support member and a monolayer of said protein layer.

12. The kit of claim 11 , wherein the support member is a petri dish.

13. Contacting an antigen with the kit according to any one of claims 1 to 12; Detecting digestion marks on the surface of the kit; A method for detecting an antigen, comprising:

14. The method of claim 13, wherein the antigen comprises a cysteine ​​protease antigen, a serine protease antigen, an aspartic protease antigen, or a metalloprotease antigen.

15. The method according to claim 13, comprising adding a reaction stopper to the kit after contacting the kit according to any one of claims 1 to 12 with an antigen.

16. The method of claim 15, wherein the quenching agent is an alcohol.

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