A kind of foot-and-mouth disease type A virus SIGA antibody ELISA detection kit and its application

A detection kit and a technology for foot-and-mouth disease are applied in the field of indirect ELISA detection kits for foot-and-mouth disease type A virus sIgA antibody, and achieve the effects of less stress on animals, less labor consumption and easy operation.

Active Publication Date: 2020-06-16
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no kit that can effectively detect sIgA antibody to foot-and-mouth disease virus in the market. In view of the in-depth research on mucosal immune mechanism and mucosal immune vaccine in recent years, an ELISA kit for detecting the level of sIgA antibody to foot-and-mouth disease virus in mucosal secretions can be developed. More accurately reflect the FMD virus infection and mucosal immune status, and use this method to monitor the dynamic changes of sIgA antibody to FMD virus, which provides theoretical and experimental basis for the FMD mucosal immunity vaccine and the establishment of supporting detection methods

Method used

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  • A kind of foot-and-mouth disease type A virus SIGA antibody ELISA detection kit and its application
  • A kind of foot-and-mouth disease type A virus SIGA antibody ELISA detection kit and its application
  • A kind of foot-and-mouth disease type A virus SIGA antibody ELISA detection kit and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0053] Preparation and assembly of embodiment 1 foot-and-mouth disease type A virus sIgA antibody ELISA detection kit

[0054] (1) Preparation of the kit:

[0055] 1. The design of the screening of the dominant epitope (TB / A) of the foot-and-mouth disease type A virus:

[0056] The structural protein VP1 of foot-and-mouth disease virus is the dominant antigen of the virus. Whether it is the isolated and purified natural VP1 protein or the recombinant expression product, it can induce the body to produce protective neutralizing antibodies, which is type-specific. The full length of the foot-and-mouth disease virus VP1 gene consists of 639 nucleotides, encoding a protein with 213 amino acids, and its main epitopes are concentrated in the 140-160 amino acid and the 200-213 amino acid segment. The present invention uses DNAStar biological software to represent strain A / GDMM / 2013 (GenBank accession number: KF450794.1), A / HuBWH / 2009 (GenBank accession number: JF792355.1), A / GDMM / 20...

Embodiment 2

[0098] Sensitivity, specificity, repeatability experiment of embodiment 2 foot-and-mouth disease type A virus sIgA antibody ELISA detection kit

[0099] 1. Sensitivity experiment:

[0100] Utilize the kit prepared in Example 1 to detect 90 nasal test sub-samples, these samples are the nasal test sub-samples collected after the experimental challenge, before any immunization is carried out, the detection method is the same as that in Example 1 by calculating the positive The detection rate was used to analyze the sensitivity of the method. The results of ELISA showed that 88 of the above samples were positive and 2 were negative, and the positive detection rate was 97.78%. Illustrate that the kit of the present invention has better sensitivity, and the results are shown in Table 4.

[0101] Table 4 ELISA kit sensitivity test

[0102] Detection method Number of samples (parts) Positive number (parts) Negative number (parts) Positive rate (%) indirect EL...

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Abstract

The invention discloses a type A foot-and-mouth disease virus sIgA antibody ELISA kit and application thereof. The kit comprises an enzyme-labeled reaction plate coated with a type A foot-and-mouth disease virus broad-spectrum multi-epitope recombinant antigen, a 100x concentrated enzyme-labeled antibody, an enzyme-labeled antibody diluent, a sample diluent, a washing concentrate, a developing solution, a stop solution, a positive control sample and a negative control sample. The type A foot-and-mouth disease virus broad-spectrum multi-epitope recombinant antigen consists of dominant antigen epitopes (TB / A) of three separated representative type A foot-and-mouth disease strains of China, so that the detection sensitivity and specificity of the kit are improved, and the kit is applied to detection of infection by different type A foot-and-mouth disease viruses. The kit is suitable for detecting sIgA antibodies in mucosal secretions of three susceptible animals, i.e., pigs, cattle and sheep, and is significant for prevention and control of propagation and infection of type A foot-and-mouth disease viruses.

Description

technical field [0001] The invention relates to an indirect ELISA detection kit for foot-and-mouth disease type A virus sIgA antibody and a detection method thereof. The invention belongs to the technical field of biological detection. Background technique [0002] Foot-and-Mouth Disease (FMD) is an infectious disease caused by the Foot-and-Mouth Disease virus, characterized by vesicular disease on the mouth and hooves of sick animals. Foot-and-Mouth Disease Virus (FMDV) belongs to the Picornaviridae family, and the center of the virus is a single-stranded RNA. According to the serotype characteristics, it can be divided into 7 serotypes: A, O, C, SAT1, SAT2, SAT3 (ie South African 1, 2, 3) and Asian (Asian 1). There is almost no immune protection between them, and animals infected with one serotype of FMDV can still be infected with another serotype of FMDV to develop disease. There are more than 70 kinds of FMDV infection objects, the incubation period is 1-7 days, the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/56983G01N33/6854G01N33/6893G01N2333/09
Inventor 潘丽张中旺吕建亮罗虹王永录张永光
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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