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Indirect ELISA kit for detecting African swine fever virus antibody and application thereof

A technology of African swine fever virus and kit, which is applied to the ELISA kit for detecting antibodies in animal quarantine, and can solve the problem of low immunogenicity in the field of detection kits for detecting African swine fever virus antibodies

Inactive Publication Date: 2011-11-09
陈文刚
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Most strains of African swine fever virus are highly virulent, but their immunogenicity is low, and only a few proteins are immunogenic

Method used

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Embodiment Construction

[0022] Below in conjunction with specific embodiment the present invention is described in further detail:

[0023] Technical scheme of the present invention is as follows:

[0024] 1. Antigen preparation

[0025] 1. Expression of African swine fever P30 protein

[0026] According to the African swine fever (ASFV) P30 gene sequence in GenBank, a pair of primers were designed and synthesized, and the P30 gene fragment was amplified from the ASFV DNA by PCR method, which was cloned into the expression vector pET28b to construct the recombinant plasmid pET-P30 After sequencing verification, it was transformed into the expression host strain BL21(DE3), and its expression was induced by IPTG.

[0027] The designed primers are:

[0028] P30-1-5'-GC GGATCC TAATTTTAAAATTGAATGGAT-3'

[0029] P30-2-5'-GT CTCGAG CCCAATCAAAATTAGATAACT-3'

[0030] The underlined part is the introduced restriction site, P30-1 is the upstream amplification primer of ASFV P30 gene, and the introduced ...

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Abstract

The invention discloses an indirect ELISA kit for detecting an African swine fever virus antibody and an application thereof, and belongs to the technical field of biology. The kit adopts prokaryotic expression recombinant P30 protein as a coating antigen, and detects the antibody of African swine fever virus in porcine serum based on the indirect ELISA principle. The coating antigen in a 96-well plate of the kit is prokaryotic expression recombinant P30 protein which has good antigenicity. The enzyme-linked immunoassay kit provided by the invention comprises a 96-well plate coated with P30 protein, a positive control, a negative control, a horseradish peroxidase-labeled rabbit anti-porcine IgG polyclonal antibody, a concentrated washing liquid, a serum diluent, a TMB substrate, and a terminating liquid. The kit of the invention is applicable to the screening of large quantities of samples, and main reagents in the kit are provided in a form of operating fluid which is convenient for use.

Description

technical field [0001] The invention is a detection kit for detecting African swine fever virus (ASFV) antibody, and belongs to the field of biotechnology. Specifically, the present invention relates to an ELISA kit for detecting antibodies in animal quarantine and its application. Background technique [0002] African swine fever (ASF) is an acute, febrile, highly contagious infectious disease of pigs caused by African swine fever virus (ASFV). It is characterized by a short course of disease and a high fatality rate, which can be as high as 100%. The clinical symptoms and pathological changes are similar to acute swine fever, and it is easy to be misdiagnosed during diagnosis. (William, Hess Adv.African swine fever: a reassessment[J].Vet Sci Comp Med, 1981, 25:39~69) The World Organization for Animals (OIE) listed it as a class A disease, and our country stipulates that it is a class I animal disease, which is subject to All countries in the world attach great importance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543C07K16/42
Inventor 陈文刚董志珍王涛
Owner 陈文刚
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