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ELISA detection kit for foot-and-mouth disease O-type virus sIgA antibody and application thereof

A detection kit, a technology for foot-and-mouth disease, applied in the directions of viruses/phages, applications, viruses, etc., to achieve the effects of less stress on animals, less time required, and less labor consumption

Active Publication Date: 2019-01-18
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

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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  • ELISA detection kit for foot-and-mouth disease O-type virus sIgA antibody and application thereof
  • ELISA detection kit for foot-and-mouth disease O-type virus sIgA antibody and application thereof
  • ELISA detection kit for foot-and-mouth disease O-type virus sIgA antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Preparation and assembly of embodiment 1 foot-and-mouth disease type O virus sIgA antibody ELISA detection kit

[0053] (1) Preparation of the kit:

[0054] 1. The design of the main neutralizing epitope (EB / O) of the representative strain of foot-and-mouth disease O-type virus:

[0055] 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 O / Akesu / 58 (NCBI accession number: AF511039.1), O / TAW / 81 / 97 (NCBI accession number: AJ296321.1) o...

Embodiment 2

[0091] Sensitivity and specificity experiment of embodiment 2 foot-and-mouth disease type O virus sIgA antibody ELISA detection kit

[0092] 1. Sensitivity experiment:

[0093] Using the kit prepared in Example 1 to detect 100 nasal test sub-samples, these samples were collected after the experimental challenge of O / Akesu / 58, O / TAW / 81 / 97, O / Tibet / 99, and O / Mya98 respectively The nose test sub-samples of diseased pigs, 25 cases of each virus sample, did not carry out any immunization before the virus challenge, and the detection method was the same as that in Example 1. The sensitivity of the method was analyzed by calculating the positive detection rate. The results of ELISA showed that 93 samples were positive and 7 samples were negative, and the positive detection rate was 93%. It shows that the kit of the present invention has good sensitivity and can detect the infection of different foot-and-mouth disease O-type viruses. The results are shown in Table 3.

[0094] Tabl...

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Abstract

The invention discloses an ELISA detection kit for a foot-and-mouth disease O-type virus sIgA antibody and an application thereof. The kit comprises a foot-and-mouth disease O-type virus broad spectrum multi-epitope recombinant antigen coated enzyme labeled reaction plate, a 100*concentrated enzyme labeled antibody, an enzyme labeled antibody diluent, a sample diluent, a concentrated cleaning solution, a developing solution, a stop solution, a positive control sample and a negative control sample. The foot-and-mouth disease O-type virus broad spectrum multi-epitope recombinant antigen is composed of main neutral epitopes of Cathay, PanAsia and Burmese 98 three pedigrees of foot-and-mouth disease O-type virus typical strains, so that the sensitivity and specificity of kit detection can be promoted and the kit is suitable for detection of different foot-and-mouth disease O-type virus infections. The kit provided by the invention is suitable for detection of sIgA antibodies in easily infected animal mucosal secretion of pig, cattle and sheep and has a great significance in preventing and controlling the spreading and infection of foot-and-mouth disease O-type virus.

Description

technical field [0001] The invention relates to an indirect ELISA detection kit for foot-and-mouth disease type O 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. The MDV nucleic acid molecule is a single-strand positive-strand RNA molecule. The total length...

Claims

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Application Information

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IPC IPC(8): C07K14/09C12N15/42G01N33/569
CPCC07K14/005C12N2770/32122G01N33/56983G01N2333/09G01N2469/20
Inventor 潘丽吕建亮张中旺罗虹王永录张永光
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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