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Competitive ELISA kit for detecting african swine fever virus antibody and purposes thereof

An African swine fever virus and kit technology, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of low immunogenicity and achieve the effect of strong practicability and reliable effect.

Inactive Publication Date: 2013-01-30
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Description
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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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  • Competitive ELISA kit for detecting african swine fever virus antibody and purposes thereof
  • Competitive ELISA kit for detecting african swine fever virus antibody and purposes thereof

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Embodiment Construction

[0022] The hybridoma cell line of the monoclonal antibody against African swine fever virus is preserved in CGMCC No. 3771 of the General Microbiology Center of the China Microbiological Culture Collection Management Committee, the preservation date: 2010-04-22, and the classification name: hybridoma cell line.

[0023] Below in conjunction with specific embodiment the present invention is described in further detail:

[0024] Technical scheme of the present invention is as follows:

[0025] 1. Establishment of cell lines

[0026] 1. Antigen Preparation

[0027] a) Expression of African swine fever P30 protein

[0028] 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...

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Abstract

The invention discloses a competitive ELISA kit for detecting african swine fever virus antibody and the purposes thereof, which belong to the biological technical field. The kit adopts prokaryotic expression recombinant P30 protein as coating antigen, and detects the african swine fever virus antibody in pig serum according to a competitive ELISA principle. The coating antigen in a 96 pore plate in the kit is the prokaryotic expression recombinant P30 protein, and has good antigenicity. The ELISA kit comprises the 96 pore plate coated by the P30 protein, and monoclonal antibody, concentrated washing liquid, serum dilution, TMB substrate and stopping solution of positive control, negative control and horseradish peroxidase labeling. The kit can be used for screening a large batch of samples, and the main reagent in the kit is provided in the form of working solution, so that the use is convenient.

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 Patents(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/543G01N33/535
Inventor 董志珍赵祥平肖妍侯艳梅王涛张瑞栾慎顺
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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