Detection kit for rapidly detecting chicken, duck and pork mixed in beef and mutton

Abstract

The utility model discloses a detection kit for rapidly detecting chicken, duck and pork mixed in beef and mutton. The detection kit comprises a detection card and a plastic card, the detection card is formed by sequentially lining absorbent paper and a sample pad below a nitrocellulose membrane, cutting the nitrocellulose membrane into test strips, and packaging the test strips in the plastic card; due to the fact that specific species antibodies of chickens, ducks and pigs are preset in the detection kit, chicken, duck or pork components which are as low as 3% and mixed in milligram-level samples can be sensitively detected at a time through antigen-antibody reaction.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the field of food safety's rapid detection, specifically relates to a rapid detection reagent box of mixed chicken, duck and pork in beef and mutton. BACKGROUND

[0002] With the improvement of living standards, the per capita meat consumption in China is also growing, among which the consumption growth of higher value beef and mutton is very outstanding compared with other poultry meat. Under the driving of illegal interests, the adulteration of beef and mutton food also occurs from time to time. This behavior not only seriously damages the interests of consumers, but also seriously damages the market order, and ultimately will seriously affect the long-term development of the industry economy. Therefore, whether it is the general public consumers or the industry administrative supervision department pays great attention to the authenticity and safety of beef and mutton food, and the inspection and identification work of other poultry source adulteration components in beef and mutton has important practical significance for ordinary consumers, and has universal social significance for the accurate law enforcement of government supervision departments.

[0003] Traditionally, many laboratory molecular biology techniques can be used to detect other poultry source components in beef and mutton products, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), mass spectrometry and infrared spectroscopy. But the above methods need to rely on special equipment and professional technicians, and the results can be obtained under laboratory conditions after a relatively long detection time. In order to more effectively protect consumers and maintain market order, it is urgent to develop a simple operation, sensitive and rapid, low-cost detection method to rapidly detect and identify other poultry source components adulterated in beef and mutton products. UTILITY MODEL CONTENT

[0004] The utility model aims at providing a kind of detection method of simple operation, sensitive and rapid, low-cost, rapidly detects and identifies other poultry source components adulterated in beef and mutton products, to solve the on-site identification problem of other poultry source components adulterated in beef and mutton products. It has significant economic benefit and social benefit. To achieve the above technical purpose, the utility model adopts the following technical scheme:

[0005] The application discloses a detection kit for rapidly detecting chicken, duck and pork mixed in beef and mutton, which comprises a detection card and a plastic card; the detection card is sequentially lined with an absorbent paper and a sample pad under a nitrocellulose membrane, cut into a test paper strip and packaged in the plastic card; the nitrocellulose membrane is provided with a quality control C line coated with rabbit anti-sheep skeletal muscle calponin T monoclonal antibody and rabbit anti-cattle skeletal muscle calponin I antibody, a detection TC line coated with gold-labeled chicken IgY monoclonal antibody, a detection TD line coated with gold-labeled duck skeletal muscle calponin I monoclonal antibody and a detection TP line coated with gold-labeled pig skeletal muscle calponin I monoclonal antibody; and the plastic card is provided with a sample loading hole.

[0006] According to the specific implementation of the application, the sample loading hole is located at the packaging portion of the plastic card and penetrates through the packaging portion, and the bottom of the sample loading hole contacts the absorbent paper; and the packaging portion is located at both sides of the plastic card.

[0007] According to the specific implementation of the application, the sample loading hole is in the shape of an inverted cone.

[0008] According to the specific implementation of the application, the upper part of the part of the absorbent paper not being packaged is further attached with a transparent film.

[0009] According to the specific implementation of the application, the colloidal gold solution is prepared by using trisodium citrate, and the gold-labeled antibody is prepared by using a freeze vacuum drying method.

[0010] Compared with the prior art, the application has the beneficial effects that:

[0011] The detection kit is provided with specific species antibodies of chicken, duck and pig, and can sensitively detect, by antigen-antibody reaction, chicken, duck or pork components mixed in milligram-level samples by one time as low as 3%; the nitrocellulose membrane is sequentially lined with an absorbent paper and a sample pad under the nitrocellulose membrane, cut into a test paper strip and packaged in the plastic card, so that the detection kit has a simple and reliable structure and can be manufactured by a standardized process; the inverted-cone-shaped sample loading hole reduces liquid residue and improves sample penetration uniformity; the transparent film covers the part of the absorbent paper not being packaged, so as to ensure that the color development result is clear and visible; the colloidal gold-labeled antibody is combined with the freeze drying process, and has no cross reaction with other animal-derived components; the detection process can adopt a standardized optimized process, and the detection kit has the advantages of convenient sampling, small sampling amount, simple operation, clear result, short time consumption, high sensitivity and the like, and can provide an objective basis for ordinary users to accurately and rapidly detect on site. BRIEF DESCRIPTION OF DRAWINGS

[0012] Figure 1 is a structure side sectional view of the detection card for detecting chicken, duck and / or pork mixed in beef and mutton according to the application;

[0013] Figure 2 is the sample detected by the utility model does not contain beef and mutton component, or the failure result schematic diagram of detection;

[0014] Figure 3 is the sample detected by the utility model does not contain beef and mutton component, but contains chicken, duck and pork result schematic diagram;

[0015] Figure 4 is the sample detected by the utility model contains beef and mutton component, and does not contain chicken, duck and pork result schematic diagram;

[0016] Figure 5 is the sample detected by the utility model contains beef and mutton component, but contains chicken result schematic diagram;

[0017] Figure 6 is the sample detected by the utility model contains beef and mutton component, but contains chicken and duck result schematic diagram;

[0018] Figure 7 is the sample detected by the utility model contains beef and mutton component, but contains chicken, duck and pork result schematic diagram.

[0019] In the figure: 1, detection card;2, plastic card;11, nitrocellulose membrane;12, water absorption paper;13, sample pad;14, transparent film;21, sample hole;22, packaging part. DETAILED DESCRIPTION

[0020] The technical solutions in the embodiments of the utility model will be clearly and completely described below with reference to the drawings in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by those skilled in the art without creative labor belong to the scope of protection of the utility model.

[0021] Antibody preparation

[0022] In the present study, purified chicken IgY was used to prepare oil emulsion immunogen to immunize mice, and the spleen cells of mice with high antibody titer were fused with myeloma cells. The monoclonal antibody in the supernatant of hybridoma cells was detected by enzyme-linked immunosorbent assay (ELISA), and the positive clones were subcloned and repeatedly screened several times. Finally, a hybridoma cell strain capable of stably secreting anti-chicken IgY was obtained. The hybridoma cell strain can secrete high-titer antibodies after repeated freezing, recovery and multiple passages, and the antibody titer remains above 1:1000. The cross-reactivity of the monoclonal antibody with duck and pig serum was determined by enzyme-linked immunosorbent assay (ELISA), and the results showed that there was no cross-reactivity. The obtained antibody has the characteristics of high affinity and strong specificity.

[0023] In the same way, duck and pig skeletal muscle troponin I (sTnI) and bovine skeletal muscle troponin I (sTnI) and goat skeletal muscle troponin T (sTnT) monoclonal antibodies were prepared.

[0024] Test card preparation

[0025] The colloidal gold solution was prepared by using trisodium citrate, and the gold-labeled antibody was prepared by using the method of freeze-drying. The gold-labeled chicken IgY monoclonal antibody was coated on the test line (TC line) of the nitrocellulose membrane (NC membrane), the gold-labeled duck skeletal muscle troponin I (sTnI) monoclonal antibody was coated on the test line (TD line) of the nitrocellulose membrane (NC membrane), and the gold-labeled pig skeletal muscle troponin I (sTnI) monoclonal antibody was coated on the test line (TP line) of the nitrocellulose membrane (NC membrane). The rabbit anti-goat skeletal muscle troponin T (sTnT) monoclonal antibody and the rabbit anti-bovine skeletal muscle troponin I (sTnI) antibody were coated on the control line (C line) of the nitrocellulose membrane (NC membrane). The nitrocellulose membrane was sequentially lined with an absorbent paper and a sample pad, cut into a test strip, and packaged in a plastic card.

[0026] A rapid detection kit for detecting chicken, duck and pig meat mixed in beef and mutton was formed, which comprises a detection card 1 and a plastic card 2. The detection card 1 is sequentially lined with an absorbent paper 12 and a sample pad 13 under a nitrocellulose membrane 11, cut into a test strip, and packaged in the plastic card 2. The nitrocellulose membrane 11 has a control C line coated with a rabbit anti-goat skeletal muscle troponin T monoclonal antibody and a rabbit anti-bovine skeletal muscle troponin I antibody, a test TC line coated with a gold-labeled chicken IgY monoclonal antibody, a test TD line coated with a gold-labeled duck skeletal muscle troponin I monoclonal antibody, and a test TP line coated with a gold-labeled pig skeletal muscle troponin I monoclonal antibody. The plastic card 2 has a sample hole 21.

[0027] The sample hole 21 is located in and penetrates through the packaging part 22 of the plastic card 2, and the bottom of the sample hole 21 contacts the absorbent paper 12. The packaging part 22 is located on both sides of the plastic card 2. The sample hole 21 is in the shape of an inverted cone. The upper part of the un-packaged part of the absorbent paper 12 is additionally attached with a transparent film 14.

[0028] Sample processing

[0029] Method one: Take out the cotton swab with the extraction solution reserved in the kit, repeatedly smear on the surface of the sample to be tested, and then put the cotton swab back into the plastic tube containing the extraction solution, tighten the tube cap, and shake the plastic tube for 10-20 seconds to fully mix.

[0030] Method two: Put 0.1g (the size of soybean) of the sample to be detected into the extraction solution in the plastic tube directly with appropriate tools, shake the plastic tube for 10-20 seconds to make the extraction solution contact the sample sufficiently, and dissolve the sample in the solution as much as possible.

[0031] Sample detection

[0032] Place the detection card in the room temperature environment, dip the extraction solution 1-2 drops with a cotton swab, drop into the sample hole of the detection card, read within 5-10 minutes, and determine the result according to the result shown in the figure.

[0033] C line does not appear, which indicates that the sample to be detected does not contain beef and / or mutton components, or the detection operation is incorrect.

[0034] C line does not appear, and only TC, TD and TP lines appear alone or simultaneously, which indicates that the sample to be detected does not contain beef and / or mutton components, but contains chicken, duck and / or pork components.

[0035] C line appears, which indicates that the sample to be detected contains beef and / or mutton components, and the detection operation is correct.

[0036] C line and TC line appear simultaneously, which indicates that the sample to be detected contains beef and / or mutton components, and contains chicken components.

[0037] C line and TC and TD lines appear simultaneously, which indicates that the sample to be detected contains beef and / or mutton components, and contains chicken and duck components.

[0038] C line and TC, TD and TP lines appear simultaneously, which indicates that the sample to be detected contains beef and / or mutton components, and contains chicken, duck and pork components.

[0039] The method has a detection limit of 3% for chicken mixed in beef and mutton, has good repeatability and specificity, other animal tissues have no interference, and the detection time only needs 15 minutes. The method is simple in operation, has high sensitivity and specificity, and is suitable for on-site rapid screening and detection of chicken mixed in beef and mutton.

[0040] The kit adopts bovine troponin I (TnI) and sheep skeletal muscle troponin T (sTnT) monoclonal antibody as a positive control, and can detect the presence of beef or mutton components.

[0041] The above merely describes a preferred specific implementation manner of the utility model, but the protection scope of the utility model is not limited to this, any skilled person in the technical field according to the technical scheme and the utility model concept of the utility model within the technical range disclosed by the utility model makes equivalent replacement or change, should be covered in the protection scope of the utility model.

Claims

1. A rapid detection kit for detecting chicken, duck and pork mixed in beef and mutton, characterized in that, The application relates to a detection card (1) and a plastic card (2); the detection card (1) is packaged in the plastic card (2) and is composed of a nitrocellulose membrane (11), a water absorption paper (12) and a sample pad (13) from bottom to top; the nitrocellulose membrane (11) is provided with a quality control C line coated with rabbit anti-sheep skeletal myocardin T monoclonal antibody and rabbit anti-cattle skeletal myocardin I antibody, a detection TC line coated with gold-labeled chicken IgY monoclonal antibody, a detection TD line coated with gold-labeled duck skeletal myocardin I monoclonal antibody and a detection TP line coated with gold-labeled pig skeletal myocardin I monoclonal antibody; the plastic card (2) is provided with a sample hole (21). The sample hole (21) is located in and penetrates through a packaging part (22) of the plastic card (2), and the bottom of the sample hole (21) contacts the water absorption paper (12); the packaging part (22) is located on both sides of the plastic card (2).

2. The rapid detection kit for detecting chicken, duck and pork mixed in beef and mutton according to claim 1, characterized in that, The sample hole (21) is in the shape of an inverted cone.

3. The test kit for rapid detection of chicken, duck and pork mixed in beef and mutton according to claim 2, characterized in that, The upper part of the water absorption paper (12) is additionally attached with a transparent film (14).

4. The test kit for rapid detection of chicken, duck and pork mixed in beef and mutton according to claim 1, wherein Colloidal gold solution is prepared by using trisodium citrate, and gold-labeled antibody is prepared by using a freeze vacuum drying method.

5. The rapid detection kit for detecting chicken, duck and pork mixed in beef and mutton according to claim 1, characterized in that, ​

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