Foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit

A foot-and-mouth disease virus and structural protein technology, which is applied in the field of the foot-and-mouth disease virus structural protein antibody enzyme-linked immunoassay detection kit and the foot-and-mouth disease virus structural protein VP1 antibody enzyme-linked immune detection kit, can solve the problems of international trade blockade, restriction, economic loss and the like , to achieve the effect of high sensitivity, reduced loss and good repeatability

Active Publication Date: 2013-11-27
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, it will trigger the market circulation of animals and animal products in endemic countries or regions, and greatly blo

Method used

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  • Foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit
  • Foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit
  • Foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit

Examples

Experimental program
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Example Embodiment

[0046] Example 1. Preparation of Enzyme-linked Immunoassay Kit for Coating Antigen of Foot-and-Mouth Disease Virus Structural Protein Antibody

[0047] According to the different epidemic strains of foot-and-mouth disease in China, this experiment uses bioinformatics methods to accurately analyze multiple subtypes of foot-and-mouth disease virus O VP1 structural protein, screen out suitable peptides, and synthesize them with an automatic peptide synthesizer. The two peptides designed by the strain sequence are shown in sequence 1 and sequence 2 in the sequence table respectively. They are made into newer and more complete coating antigens with a purity of about 90%, which can adapt to the continuous mutation characteristics of foot-and-mouth disease virus and improve Detection rate of antibody positive. The polypeptide synthesis method can be a conventional method. The present invention uses the following methods to synthesize the two polypeptides of the present invention as the ...

Example Embodiment

[0078] Example 2. Preparation of an enzyme-linked immunoassay kit for foot-and-mouth disease virus structural protein antibody

[0079] Foot-and-mouth disease virus structural protein antibody enzyme-linked immunoassay kit includes:

[0080] (1) 96-well removable polystyrene enzyme-linked reaction plate coated with foot-and-mouth disease virus antigen; 2×96 wells.

[0081] (2) Horseradish peroxidase labeled rabbit anti-pig IgG polyclonal antibody (purchased from sigma company, article number A5670), 2 bottles (12ml each).

[0082] (3) Positive control serum: using the artificially synthesized coating antigen in Example 1 as the immunogen, the hyperimmune serum prepared by immunizing 25-35-day-old SPF pigs with 1000U / ml streptomyces And 1000U / ml penicillin, sterilized through 0.2μm filter membrane, as the positive control serum of the kit. 1 tube (1ml).

[0083] (4) Negative control serum: normal pig serum without foot-and-mouth disease antibodies, 1000U / ml streptomycin and 1000U / ml pe...

Example Embodiment

[0090] Example 3. Compliance rate test of foot-and-mouth disease virus structural protein antibody enzyme-linked immunoassay kit

[0091] 1. How to use foot-and-mouth disease virus structural protein antibody enzyme-linked immunoassay kit

[0092] 1. Equilibrium: Take out the kit stored at 4°C and equilibrate it to room temperature for later use; mix the liquid reagent before use.

[0093] 2. Liquid preparation: dilute the concentrated washing solution 20 times with distilled water or deionized water to obtain washing buffer;

[0094] 3. Setting: Leave 1 blank well, 3 negative control wells, and 2 positive control wells, and the rest are sample wells to be tested.

[0095] 4. Pre-dilution of the test specimen: Use the sample diluent to dilute the test sample serum, the negative control serum, and the positive control serum at a ratio of 1:21; add 100μl of sample diluent to each well of the dilution plate, and add 5μl of the sample to each well. Test samples.

[0096] 5. Adding samples: ...

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Abstract

The invention discloses a foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit. The kit comprises an enzyme-linked reaction plate coated by foot and mouth disease virus structural protein VP1 antigenic epitope polypeptides, and an enzyme-labeled antibody. The foot and mouth disease virus structural protein VP1 antigenic epitope polypeptides are a polypeptide represented by the sequence 1 in the sequence list and a polypeptide represented by the sequence 2 in the sequence list. According to the kit, the reaction plate coated by chemically synthesized VP1 antigenic peptide, antigen dose is low, sensitivity is high, and specificity is high. With the kit, whether foot and mouth disease virus infection exists can be highly efficiently detected. The kit provided by the invention has good specificity, high sensitivity, high efficiency, and good market prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, the invention relates to an enzyme-linked immunosorbent detection kit for an antibody to the structural protein of foot-and-mouth disease virus, in particular to an enzyme-linked immunosorbent detection kit for an antibody to the structural protein VP1 of foot-and-mouth disease virus. Background technique [0002] Foot-and-Mouth Disease (FMD) is a global epidemic infectious disease that is widely distributed in the world and spread internationally. The International Bureau of Veterinary Medicine (OIE) listed foot-and-mouth disease as the first of Class A infectious diseases in the revised classification of animal infectious diseases in 1984. Foot-and-mouth disease is a highly contagious zoonotic disease of artiodactyls caused by Foot-and-Mouth Disease Virus (FMDV) infection. It is characterized by acute onset, rapid spread, and high infection rate, resulting in a rapid decline in ...

Claims

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Application Information

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IPC IPC(8): C07K14/09G01N33/68G01N33/569G01N33/543G01N33/545
CPCC07K14/09G01N33/56983G01N2333/09
Inventor 董春娜齐鹏肖进栗利芳王楠巴利民宋芳张蕾周强郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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