Immunochromato kit

Abstract

To provide an immunochromato test kit which, in a detection method for parasites in solid food, particularly seafood, using an immuno-chromatography method, enables simple processing of a sample without using equipment (such as a stomacher or homogenizer) employed in pretreatment for general food inspection to grind and homogenize the sample, and which allows more efficient extraction and detection of a target substance (antigen protein).SOLUTION: An immunochromato kit comprises a net used for grinding a sample, a grinding bag for containing the sample for performing grinding treatment, an extraction solution for extracting a target antigen protein from the sample, and an immunochromato test strip, where the net satisfies the following condition (1-1) or (1-2) and condition (2): (1-1) an area of a mesh is greater than or equal to 0.4 mm2 and less than 23.5 mm2, and a ratio of the short side to the long side (short side / long side) of the mesh is 1 to 0.4; (1-2) an area of the mesh is greater than or equal to 23.5 mm2, and a width of a twisted yarn is greater than or equal to 1.2 mm and less than 3.0 mm; and (2) a thickness of the net is greater than or equal to 0.2 mm and less than 3.5 mm.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an immunochromatography kit. [Background technology]

[0002] Health hazards caused by parasites in seafood include food poisoning and food allergies. The number of cases of food poisoning caused by Anisakis (a nematode) parasitizing fresh seafood is increasing every year. The larvae (hereafter referred to as Anisakis larvae) parasitize seafood such as mackerel, horse mackerel, saury, bonito, sardines, and salmon. Anisakis larvae are known to migrate from the intestines to the muscles after the infested seafood dies. When fresh seafood infested with Anisakis larvae is eaten raw (or insufficiently cooked or frozen), the larvae penetrate the stomach or intestinal wall, causing food poisoning (anisakiasis). The number of cases of Anisakis food poisoning has been on the rise since 2018, and it is speculated that this is due to factors such as continued high seawater temperatures. Currently, there are 1,061 cases of food poisoning per year, of which 328 (31%) are caused by Anisakis, followed by Campylobacter with 286 (27%) and Norovirus with 212 (20%). Anisakis allergy is an allergy caused by the protein of Anisakis. Due to the high domestic consumption of seafood in Japan, seafood is the second most common cause of food allergies after eggs. However, it has become clear that the overwhelming majority of cases are caused by the Anisakis larvae that parasitize seafood, rather than the seafood itself. Even if there are no live Anisakis when eating seafood, traces of its presence, such as dead Anisakis, body fluids, secretions, or excrement, can cause an allergic reaction. Official parasitic testing methods (notifications) are currently available for Kudoa (Ministry of Health, Labour and Welfare: Food and Drug Administration Notification No. 0427-3) and Sarcocystis (Ministry of Health, Labour and Welfare: Food and Drug Administration Notification No. 0427-4), but not for Anisakis. Currently, visual inspection and the candling method are the primary methods for testing for Anisakis. To prevent food poisoning and allergies caused by Anisakis, the general method involves visually inspecting and removing Anisakis larvae. However, the direct observation method, which involves visually inspecting the abdominal cavity, visceral surfaces, and muscles of infected seafood to detect Anisakis larvae, is inefficient. The candling method uses ultraviolet light instead of white light to improve detection efficiency, but requires expensive equipment. Furthermore, when attempting to carry out detection by genetic testing, nucleic acids must be extracted from the sample, and there is the problem that PCR conditions and settings require time and effort. Immunochromatography is sometimes used as a simple method in food testing to detect food ingredients, allergens, toxins, and parasites present in food. Immunochromatography testing of solid foods requires crushing and homogenizing the solids in a liquid. Food analysis typically involves the use of instruments such as stomachers and homogenizers to crush and homogenize samples. However, stomachers are expensive and not commonly used. Surfactants are sometimes used to elute target substances from samples, but the surfactants can cause foaming, making it difficult to recover the extract. Homogenizers can mechanically crush samples, while food processors and blenders use rotating blades to crush and homogenize samples, but they require maintenance, such as cleaning the container after each use. Furthermore, mortars and pestles used for manual sample crushing, as well as simple microtube homogenizers (which manually crush samples using a pestle in a microtube), lack blades, making it difficult to break down pieces of meat, bones, and skin. Regarding methods for detecting parasites in solid foods, particularly seafood, Patent Document 1 discloses an enzyme-linked immunosorbent assay (ELISA) for detecting antigens of Anisakis species in food extracts, but makes no mention of methods for preparing food extracts. Patent Document 2 describes an immunochromatographic method for detecting Kudoa myxosporean parasites, which are the causative agent of food poisoning caused by eating raw flounder, but extracts the parasites from flounder muscle using a microtube homogenizer. When detecting the presence of parasites or parasite-derived allergens in solid foods, particularly seafood, there is a need for a simple testing method that can easily process samples and more efficiently extract parasite proteins without using special equipment. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Special Publication 2009-542243 [Patent Document 2] Patent Publication No. 2015-127666 Summary of the Invention [Problem to be solved by the invention]

[0004] The present invention aims to provide an immunochromatographic testing kit for a method for detecting antigen proteins in solid foods using immunochromatography, which can easily process samples without using equipment (such as a stomacher or homogenizer) that is used in pretreatment for general food testing to crush and homogenize samples, and which can more efficiently extract and detect test substances (antigen proteins). [Means for solving the problem]

[0005] The inventors have discovered that an immunochromatography kit containing a net for grinding samples allows samples to be processed easily without the need for equipment (such as a stomacher or homogenizer) used in general food testing pretreatment to grind and homogenize samples, and that the test substance (antigen protein) can be extracted and detected more efficiently.

[0006] That is, the present invention is as follows. [1] An immunochromatography kit including a net (mesh) used for grinding a sample, a grinding bag for placing the sample in and grinding it, an extractant for extracting a target antigen protein from the sample, and an immunochromatography test piece, The net satisfies the following conditions (1-1) or (1-2) and (2) (1-1) Mesh area is 0.4 mm 2 Over 23.5mm 2 and the ratio of the length of the long side to the short side of the mesh (short side / long side) is 1 to 0.4, or (1-2) Mesh area is 23.5 mm 2 or more, and the width of the twisted yarn is 1.2 mm or more and less than 3.0 mm; (2) The thickness of the net is 0.2 mm or more but less than 3.5 mm; The immunochromatography kit. [2] The immunochromatography kit of [1], wherein the knitting method of the net is selected from raschel knitting, stockinette knitting, cord knitting, and plain weave. [3] Immunochromatography kits [1] and [2] for detecting parasites in seafood. [4] An immunochromatography kit for the parasite Anisakis [3]. [5][4] A method for detecting Anisakis using the immunochromatography kit. [Effects of the Invention]

[0007] By using the immunochromatography kit of the present invention, in a method for detecting antigen proteins in solid foods using immunochromatography, particularly in a method for detecting parasites in seafood, samples can be processed easily without using equipment (such as a stomacher or homogenizer) that is used in general food testing pretreatment to crush and homogenize samples, and the test substance (antigen protein) can be extracted and detected more efficiently. [Brief explanation of the drawings]

[0008] [Figure 1] This is an explanation of each part of the net. [Figure 2] FIG. 1 is a schematic diagram of an immunochromatographic test piece (test strip). DETAILED DESCRIPTION OF THE INVENTION

[0009] The present invention relates to an immunochromatography kit comprising a net (mesh) having predetermined properties used for grinding a sample, a grinding bag into which the sample is placed for grinding, an extractant for extracting a target antigen protein from the sample, and an immunochromatography test piece.

[0010] The samples of the present invention include seafood, meat, vegetables, fruits, processed eggs, dairy products, and solid foods made using these. In view of the difficulty of grinding the samples, the effects of the present invention can be more effectively achieved when the samples are collected from seafood.

[0011] In the present invention, seafood refers to fish, crustaceans, shellfish, and cephalopods. Seafood also includes edible marine mammals such as whales and dolphins. The fish meat sample refers to all edible parts of the fish, including the head, body (red meat, white meat, bones, etc.), internal organs, and tail fin. From the standpoint of workability in grinding, it is preferable to remove hard parts such as fish bones and skin, the exoskeleton of crustaceans, the shells of shellfish, and the carapace and mantle of cephalopods. However, it is also possible to sample the fish without removing them and grind the remaining parts together.

[0012] In the present invention, the net is preferably a woven or knitted fabric. Woven fabrics are fabricated by crossing two sets of threads, warp and weft, while knitted fabrics are fabricated by intertwining or knotting string-like threads. Net weaving methods can be any common woven or knitted fabric manufacturing method, including, for example, raschel knitting, stockinette knitting, cord knitting, and plain weave. A net-like structure can also be formed by welding the warp and weft threads formed by extrusion molding at their intersections (intersection welding). The principle of net formation by extrusion molding is that holes for extruding the threads are drilled in half of each mold on the contact surfaces of separate molds that are in contact with each other. By sliding against each other perpendicular to the extrusion direction, meshes or knots are formed at the points where the holes in the two mold halves meet, and the remaining areas become net threads. To achieve the effects of the present invention, knitted or woven nets are preferred, with raschel knitting and stockinette knitting being more preferred. The netting fibers may be made of either synthetic or natural fibers, but synthetic fibers are preferred because they are relatively hard when knitted or woven into a net and have a low moisture absorption rate. Typical synthetic fiber materials include polyethylene, polypropylene, polyester, nylon, and vinylon.

[0013] The net is preferably woven using single fibers or twisted yarns made up of multiple single fibers with a diameter of approximately 0.1 mm to 0.5 mm as warp, weft, or knitting yarn, and the mesh shape of the net is not limited and can be rectangular, such as square, rectangular, or rhombic (diamond, trapezoid), or circular. In the present invention, mesh size is the dimension between the inner ends of the yarns constituting the net, and is expressed as the length of the long and short sides of the mesh. For example, if the mesh is rectangular, the longer side is the long side and the shorter side is the short side. If the mesh is rhombic or parallelogram, the longer diagonal is the long side and the shorter diagonal is the short side. If the mesh is elliptical, the long axis is the long side and the short axis is the short side. If the mesh is square or circular, the long and short sides are the same length. Twisted yarn refers to a weft, warp, or knitting yarn made by twisting a bundle of multiple single fibers, and the width of the twisted yarn refers to the width of the widest weft, warp, or knitting yarn. An example of a net of the present invention is shown in Figure 1.

[0014] In the present invention, the net satisfies the following conditions (1-1) or (1-2) and (2): (1-1) Mesh area is 0.4 mm 2 Over 23.5mm 2 and the ratio of the length of the long side to the short side of the mesh (short side / long side) is 1 to 0.4, or (1-2) Mesh area is 23.5 mm 2 or more, and the width of the twisted yarn is 1.2 mm or more and less than 3.0 mm; (2) The thickness of the net is 0.2 mm or more but less than 3.5 mm. This structure prevents hard parts of the sample, such as skin and small bones, from getting caught in the mesh when the sample is ground, and the sample becomes entangled in the threads that make up the net, allowing the sample to be ground efficiently.

[0015] The mesh area is 0.45 mm 2 Over 16mm 2 More preferably, it is 0.45 mm or less. 2 Over 14mm 2 It is even more preferable that: The ratio of the length of the long side to the short side of the mesh opening (short side / long side) is more preferably 1 to 0.5, and even more preferably 0.9 to 0.6. Mesh area is 23.5mm 2 If the width is more than this, the sample tends to get caught in the mesh and become difficult to grind. Even in this case, if the width of the twisted yarn is 1.2 mm or more and less than 3.0 mm, it can be ground relatively well. The width of the twisted yarn is more preferably 1.5 mm or more and less than 2.5 mm, and even more preferably 1.5 mm or more and 2.2 mm or less. If the thickness of the net is too bulky, the sample tends to be difficult to move and grind. That is, the thickness of the net is more preferably 0.4 mm or more and less than 3.0 mm, even more preferably 0.5 mm or more and less than 2.5 mm, and even more preferably 0.5 mm or more and less than 1.5 mm.

[0016] In the present invention, the grinding bag into which the sample is placed and ground may be made of any strong and durable material, but is preferably made of transparent plastic so that the grinding state can be visually confirmed, and more preferably made of nylon / polyethylene film. The size of the grinding bag should be slightly larger than the size of the net to be used. For example, a net cut to about 50mm x 50mm can be placed in a 120mm x 70mm nylon bag with a zipper. During grinding, the bag must be sealed to prevent the extract from leaking out. A sealer can be used to seal the opening, or if the bag already has a zipper, the sample can be taken in and out and the extract can be collected efficiently.

[0017] In the present invention, the extraction solution used to extract the target antigen protein from a sample may have a composition suitable for the antigen protein to be detected. However, a buffer with high buffer capacity at a neutral pH is generally used to prevent denaturation of the antigen protein. For example, phosphate buffer, phosphate-buffered saline (PBS), Tris·HCl buffer, citrate buffer, HEPES buffer, etc. can be used. Furthermore, a surfactant may be added to the buffer to increase the solubility of the antigen protein. Any surfactant generally used in the food or biochemistry fields can be used, and the surfactant can be selected depending on the properties of the antigen protein.

[0018] The main components of the extract that can be used in the present invention are preferably a buffer solution and a surfactant. From the viewpoint of improving detection sensitivity and reducing false positive reactions, the extract in the present invention is preferably a potassium phosphate buffer solution, a sodium phosphate buffer solution, or PBS, and the concentration thereof is preferably 20 mM to 50 mM, and the pH is preferably in the range of 7.4 to 8.0.

[0019] The surfactant is preferably a nonionic surfactant, such as Tween 20, Triton X-100, or IGEPAL CA-630, and its concentration is preferably in the range of 0.05 (v / v)% to 1.6 (v / v)%.

[0020] The extract solution of the present invention may contain a blocking agent. A hydrophilic polymer can be added to the extract solution as a blocking agent to suppress nonspecific binding of hydrophobic analyte molecules (such as proteins contained in the sample) on the immunochromatographic test piece. Examples of hydrophilic polymers include polyvinylpyrrolidone K30 (PVP-K30) and polyvinyl alcohol (PVA), with PVP-K30 being more preferred, and its concentration preferably being 0.5 (v / v)% or less.

[0021] The extract of the present invention may further contain a reducing agent. The reducing agent can prevent a decrease in extraction efficiency, so it is preferable to add a reducing agent to the extract. Considering safety and storage stability, examples of reducing agents that can be used include α-tocopherol, β-tocopherol, γ-tocopherol, σ-tocopherol, BHA (butylhydroxyanisole), BHT (dibutylhydroxytoluene), L-ascorbate, and isoascorbate, which are already used in foods and cosmetics. The concentration of the reducing agent in the extract is preferably 0.005 to 7 (w / v)%, more preferably 0.005 to 1.5 (w / v)%.

[0022] The extract of the present invention may further contain a preservative, which is preferably added to the extract to prevent the invasion, growth, and proliferation of microorganisms in the buffer solution and minimize spoilage. Any preservative commonly used in foods and cosmetics can be used without any particular limitation. Specific examples include sodium benzoate, calcium propionate, sodium propionate, and Proclin 300 (Sigma-Aldrich), with Proclin 300 being preferred. The concentration of the preservative in the extract is preferably 0.01 to 0.05 (w / v)%, more preferably 0.015 to 0.035 (w / v)%.

[0023] The extract of the present invention may further contain a chelating agent. It is preferable to add a chelating agent to the extract because it has the property of capturing metal ions that promote oxidation. The chelating agent can be any component that is usually added to foods and cosmetics for the purpose of maintaining the quality, and examples of the chelating agent include EDTA (ethylenediaminetetraacetic acid), NTA (nitrilotriacetic acid), and DTPA (diethylenetriaminepentaacetic acid). The concentration of the chelating agent in the extract is preferably 0.1 to 0.7 mM, more preferably 0.5 mM.

[0024] The immunochromatography kit of the present invention includes an immunochromatography test strip. In the immunochromatography test strip included in the immunochromatography kit, an antibody against the test substance (antigen) is preferably immobilized on a solid phase, which is a support, and is included in the form of, for example, a type in which the immunochromatography membrane with the immobilized antibody and each component are laminated on a backing sheet and the upper surface of the immunochromatography test strip is covered with a sheet-like film, or a type in which the immunochromatography test strip is housed in a molded plastic container that is molded taking into consideration the position where the sample will be dropped and the position where the antibody is immobilized on the membrane, etc.

[0025] A schematic diagram of an example of an immunochromatographic test strip is shown in Figure 2. An antibody specific to the analyte is immobilized in the detection zone (10 in Figure 2) of the test strip membrane (9 in Figure 2). In addition, a detection reagent such as a dye-labeled antibody is held in the conjugate pad (8 in Figure 2). The sample is triturated in an extract solution to elute the analyte into the extract solution, and when this extract solution is dropped onto the sample pad (7 in Figure 2) of the test strip, it develops together with the detection reagent (in the direction from 7 to 12 in Figure 2), forming a complex of immobilized antibody, analyte, and dye-labeled antibody in the detection zone (10 in Figure 2). As a result, a signal appears in the detection zone, allowing the analyte to be detected.

[0026] In the present invention, the test substance (antigen) is contained in the sample and is, for example, a pathogen, a parasite, or a partial protein, protein fragment, or peptide thereof, or a chemical substance derived from the body fluids, secretions, or excretions of the pathogen or parasite. Examples of parasites include Anisakis, Kudoa, and Sarcocystis.

[0027] In the present invention, antibodies are labeled as needed for detection in immunochromatographic testing. Labeling can be performed by conventional methods. Labeling substances used for labeling include colloidal metal particles (colloidal particles of gold, silver, copper, iron, platinum, palladium, and mixtures thereof (e.g., mixtures of gold and platinum, mixtures of gold and silver, and mixtures of palladium and platinum)) and colored particles such as colored latex particles. Among colloidal metal particles, gold colloidal particles are preferred because of their ease of use.

[0028] The immunochromatography kit of the present invention includes the above-mentioned net (mesh), trituration bag, extract, and immunochromatography test strip. In one form of kit, the net and extract can be pre-sealed in a trituration bag with a zipper. Alternatively, the entire kit can be placed in a sealed, light-blocking aluminum packaging bag to prevent deterioration of reagents such as antibodies. The immunochromatography kit of the present invention may further include any components commonly used in the art. For example, it may further include an extraction or dilution buffer, a standard for the test substance, a dropper for dripping the trituration solution, a stick for grinding the sample, a desiccant, and an instruction manual explaining how to use the kit.

[0029] The immunochromatography kit of the present invention may be used to detect parasites in seafood, such as Anisakis, Kudoa, and Sarcocystis, with Anisakis being preferred.

[0030] The present invention also relates to a method for detecting anisakis using the above-mentioned immunochromatography kit. The method for detecting anisakis of the present invention is characterized by comprising extracting a test substance (antigen) from a sample using the net, trituration bag, and extraction solution of the above-mentioned immunochromatography kit, applying the extraction solution containing the extracted test substance (antigen) to the immunochromatography test piece of the above-mentioned immunochromatography kit, and detecting the test substance (antigen) from the immunochromatography test piece. The method for detecting anisakis of the present invention can be carried out according to conventional methods, except for applying the extraction solution containing the test substance (antigen) extracted using the net, trituration bag, and extraction solution of the above-mentioned immunochromatography kit to the immunochromatography test piece, and detecting the test substance (antigen) from the immunochromatography test piece.

[0031] In the method for detecting anisakis of the present invention, the method for extracting the test substance (antigen) from the sample can be carried out in accordance with conventional methods, except for using the net, grinding bag, and extraction solution of the above-mentioned immunochromatography kit. For example, the sample, extract and net are placed in a grinding bag, and the bag is rubbed with a stick to grind the material.

[0032] In the method for detecting anisakis of the present invention, the test substance (antigen) can be detected by a conventional method. For example, antibodies labeled with colored particles such as gold colloids accumulate in the detection zone of an immunochromatographic test strip, causing a colored test line to appear, which can be visually confirmed. Alternatively, the absorbance of the test line can be measured using an immunochromatographic reader. [Example]

[0033] EXAMPLES The present invention will be described in detail below with reference to examples, but the present invention is not limited to the following examples.

[0034] <Preparation of immunochromatographic test strips for detecting Anisakis> 1. Preparation of Anti-Anisakis Antibodies Anisakis larvae collected from fish were crushed in PBS to obtain an extract. The extract was then immunized in a rabbit using standard antibody production methods to obtain antiserum. The antiserum was purified using a Protein A column to obtain anti-Anisakis polyclonal antibodies. 2. Preparation of immunochromatographic test strips A portion of the anti-Anisakis polyclonal antibody used in the immunochromatographic test strip of the present invention was labeled with gold colloid particles and used as the labeled antibody. Anti-Anisakis polyclonal antibody labeled with gold colloid particles was applied to a conjugate pad (8 in Figure 2) and allowed to dry. Anti-Anisakis polyclonal antibody was immobilized as a test line (10 in Figure 2) on the upstream side of the membrane (9 in Figure 2) and simultaneously immobilized as a control line (11 in Figure 2) on the downstream side of the membrane. The test strip was assembled by attaching the antibody-immobilized membrane (9 in Figure 2) to a backing sheet (13 in Figure 2), attaching a conjugate pad (8 in Figure 2) with dried labeled antibody to the upstream side of the membrane, and then attaching a cotton sample pad (7 in Figure 2) further upstream. A cotton absorbent pad (12 in Figure 2) was attached to the downstream side of the membrane. The assembled sheet was cut to a width of 5 mm to complete the test strip.

[0035] <Consideration of net properties and measurement results> A mesh (No. 1 to 12) cut to 50 mm x 50 mm was placed in each of the grinding bags (120 mm x 70 mm) with a zipper. *1 5 mL of the net and 0.5 g of Anisakis-negative food (sea mackerel) were added, one Anisakis larva (body length: 20 mm) was added to each trituration bag, and the trituration bag zipper was closed. A roller (made of Duracon material) was used to apply uniform pressure from above the trituration bag, moving the net back and forth and left and right five times to triturate the food. After trituration, 150 μL of the extract was collected and dropped onto the sample pad of the test strip. After 30 minutes, the absorbance of the test line was measured using an immunochromatographic reader (Hamamatsu Photonics, C10066-10). A value of 5 mABS or higher was considered positive, and a value of less than 5 mABS was considered negative. The characteristics of the net and the measurement results are shown in Tables 1 and 2. *1 The extraction solution was 0.1 (v / v)% Tween 20 / 40 mM potassium phosphate buffer (pH 7.4). Table 1 TIFF2025136237000001.tif135156 *2 Net No. 2 is the length of the long axis, and No. 6 is the length of the longer diagonal *3 Net No. 2 is the length of the minor axis, and No. 6 is the length of the shorter diagonal. Table 2 TIFF2025136237000002.tif130156 *4 Net No. 8 and No. 12 are the length of the longer diagonal *5 Net No. 8 and No. 12 are the length of the shorter diagonal In Examples 1 and 2, the mesh was relatively fine, and it was possible to grind the sample efficiently. As the mesh area increased, grinding became more difficult, and the measured value of the immunochromatographic reader tended to decrease (Example 3). 2 In Comparative Example 1, the sample remained as lumpy meat chunks and was not ground, and the immunochromatographic reader measurement value was less than 5 mABS, meaning that the Anisakis antigen could not be detected. In Example 4, the mesh area is 56.8 mm 2 Although the net was large, the width of the twisted threads was 2.2 mm, which made it easy for the fish bones and skin to get tangled and be crushed, which is thought to be why the antigenic protein of Anisakis larvae was eluted into the extract. It was observed that when the mesh is coarse, the sample tends to clump and become difficult to crush. Comparative Example 2 was an anti-slip mat with continuous diamond-shaped openings formed by extrusion molding, and although the size of the openings was relatively small, it was not made of woven twisted yarn, so the sample did not get caught on the yarn and was difficult to grind, which is thought to be why it could not be detected. In Examples 5 and 6, the samples became clumpy and paste-like and were difficult to grind, but detection was possible. In Comparative Example 3, the yarn was knitted in a three-dimensional, napped shape and was thick and bulky, so the sample did not move easily and was difficult to grind, making it difficult to detect. In Comparative Examples 4 and 5, the sample was not woven, so it became paste-like and could not be ground. In Example 7, the mesh size was relatively large and the sample got caught in the mesh and was difficult to grind, but detection was possible. From the above, it was found that the effects of the invention are more easily achieved when the grinding net included in the immunochromatography kit of the present invention is made of woven synthetic fibers with a relatively fine mesh and a relatively wide twisted yarn. [Explanation of symbols]

[0036] 1 Single fiber 2. Twisted Yarn 3. Width of twisted yarn 4 Mesh size Longitudinal 5 Mesh size Short end 6 Mesh 7 Sample pad (sample drop area) 8 Conjugate pad (dye-labeled antibody holding area) 9. Membrane (carrier) 10 Test line (detection area) 11 Control Line 12 Absorbent pad (sample liquid absorption part) 13 Backing sheet (mount)

Claims

1. An immunochromatography kit including a net (mesh) used for grinding a sample, a grinding bag for placing the sample in and grinding it, an extracting solution for extracting a target antigen protein from the sample, and an immunochromatography test piece, The net satisfies the following conditions (1-1) or (1-2) and (2): (1-1) Mesh area is 0.4 mm 2 Over 23.5mm 2 or less, and the ratio of the length of the long side to the short side of the mesh (short side / long side) is 1 to 0.4; or (1-2) Mesh area is 23.5 mm 2 or more, and the width of the twisted yarn is 1.2 mm or more and less than 3.0 mm; (2) The thickness of the net is 0.2 mm or more and less than 3.5 mm. The immunochromatography kit.

2. 2. The immunochromatography kit according to claim 1, wherein the knitting method of the net is selected from the group consisting of raschel knitting, stockinette knitting, cord knitting, and plain weave.

3. The immunochromatography kit according to claim 1 or 2, for detecting parasites in fish and shellfish.

4. The immunochromatography kit according to claim 3, wherein the parasite is Anisakis.

5. A method for detecting Anisakis, which uses the immunochromatography kit according to claim 4.

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