Competitive Alpha LISA (linked immuno sorbent assay) detection kit for classical swine fever virus (CSFV) antibody and detection method thereof

A detection kit and swine fever virus technology are applied in the field of immunological detection of swine fever virus antibodies, which can solve the problems of large differences in specificity and high requirements, and achieve short detection time, strong anti-quenching ability, and low serum dosage. Effect

Inactive Publication Date: 2014-01-08
INSPECTION & QUARANTINE TECH CENT OF CHONGQING ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
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AI Technical Summary

Problems solved by technology

The existing detection methods mainly include virus isolation and identification, immunoperoxidase technology, immunocolloidal gold technology, immunofluorescent antibody staining technology, ELISA, PCR and real time PCR, etc., each has its advantages and disadvantages, and the specificity varies greatly
At the same time, the method has higher requirements on samples and personnel
However, there are no reports on kits and detection methods for detecting CSFV antibodies using AlphaLISA technology at present

Method used

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  • Competitive Alpha LISA (linked immuno sorbent assay) detection kit for classical swine fever virus (CSFV) antibody and detection method thereof
  • Competitive Alpha LISA (linked immuno sorbent assay) detection kit for classical swine fever virus (CSFV) antibody and detection method thereof
  • Competitive Alpha LISA (linked immuno sorbent assay) detection kit for classical swine fever virus (CSFV) antibody and detection method thereof

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Embodiment Construction

[0023] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0024] The attenuated CSFV vaccine strain used in the following implementation is a commercially available product, which is stored in the Inspection and Quarantine Technology Center of Chongqing Entry-Exit Inspection and Quarantine Bureau.

[0025] 1. Construction and identification of CSFV E2 recombinant protein prokaryotic expression plasmid

[0026] 1. Target fragment amplification

[0027] 1.1, RT-PCR amplification

[0028] Using the genome of attenuated CSFV vaccine strain CSFV (GenBank No.: NC_002657.1) as a template, carry out reverse transcription polymerase chain reaction (RT-PCR) to amplify the desired fragment CSFV E2 gene, i...

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Abstract

The invention discloses a competitive Alpha LISA (linked immuno sorbent assay) detection kit for a classical swine fever virus (CSFV) antibody and a detection method thereof. The detection kit comprises donor microspheres, receptor microspheres, a swine fever virus E2 protein monoclonal antibody and an E2 protein antigen with a His label. A competitive Alpha LISA detection method for the CSFV antibody is created by optimizing test reaction conditions such as the donor microspheres, the receptor microspheres, the monoclonal antibody, the antigen and serum. The kit for detecting the CSFV antibody is good in specificity, high in sensitivity, low in usage amount of the serum, low in detection cost and short in detection time, does not need to be washed and can not be influenced by hemolysis.

Description

technical field [0001] The invention relates to the immunological detection of classical swine fever virus antibody, in particular to a detection kit and detection method for the competition AlphaLISA of classical swine fever virus antibody. Background technique [0002] Classical swine fever virus (CSFV) is a member of the Flavivirus genus. Due to the high pathogenicity and high lethality of CSFV, it has caused huge losses to the breeding industry. After pigs are infected with highly pathogenic strains, they can cause infection during the incubation period. Pigs experiencing clinical symptoms of acute swine fever or subacute swine fever shed the virus in large quantities before the onset of symptoms, while pigs that have survived have obvious CSFV antibodies in their bodies, and No more detoxing. Pigs infected with milder strains are chronically infected and shed chronically or intermittently before dying. Pregnant sows can infect fetuses through the placenta, resulting ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/56983G01N33/54313G01N33/561G01N2333/183G01N2469/20
Inventor 聂福平杨俊王昱李贤良李应国肖进文
Owner INSPECTION & QUARANTINE TECH CENT OF CHONGQING ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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