Recombination protein for detection of virus of porcine reproductive and respiratory syndrome and preparation method and application thereof

A technology for respiratory syndrome and recombinant protein, which is applied in the field of recombinant protein and its preparation for detection of porcine reproductive and respiratory syndrome virus, which can solve the problems of inability to establish a detection laboratory, high requirements for detection methods, and high detection costs

Active Publication Date: 2014-04-16
GUANGXI VETERINARY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Current PRRSV detection techniques include virus isolation and identification, virus neutralization test, immunoperoxidase monolayer test, indirect immunofluorescence antibody test, immunohistochemistry, enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction, Real-time fluorescent quantitative reverse transcription-polymerase chain reaction, in situ hybridization technology, gene chip technology, reverse transcription-loop-mediated isothermal amplification technology, etc., but ...

Method used

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  • Recombination protein for detection of virus of porcine reproductive and respiratory syndrome and preparation method and application thereof
  • Recombination protein for detection of virus of porcine reproductive and respiratory syndrome and preparation method and application thereof
  • Recombination protein for detection of virus of porcine reproductive and respiratory syndrome and preparation method and application thereof

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Experimental program
Comparison scheme
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Embodiment Construction

[0020] 1. Construction of anti-human erythrocyte H antigen single-chain antibody gene

[0021] 1. Cloning of antibody variable region genes

[0022] 1.1 Primer design

[0023] Primers were designed and synthesized based on the sequence of the monoclonal antibody variable region gene (Table 1).

[0024] Table 1 Monoclonal antibody variable region gene amplification primers

[0025]

[0026] Primers were synthesized by Beijing Biomed Biological Company. Among them, the primers VH-F and VH-R are used to amplify the antibody heavy chain variable region gene, and the primers VL-F and VL-R are used to amplify the antibody light chain variable region gene.

[0027] 1.2 Preparation of cDNA

[0028] Hybridoma cells secreting anti-human erythrocyte H antigen monoclonal antibody (1×10 6 ), using the TRIzol method to extract total cellular RNA, and then using the total RNA as a template, oligo (dT) 18 As a primer, perform reverse transcription according to the manual of reverse t...

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Abstract

The invention discloses a recombination protein for detection of virus of porcine reproductive and respiratory syndrome, which is a fusion gene code spliced by a single-chain antibody gene resisting surface H antigen of human red blood cell and a single-chain antibody gene resisting the virus of porcine reproductive and respiratory syndrome. The recombination protein has dual functions of being capable of both combining with human red blood cell without agglutinating and combining with the virus of porcine reproductive and respiratory syndrome, and has observable agglutination phenomenon under action of virus bridging. The recombination protein prepared by the method can be used for detecting virus of porcine reproductive and respiratory syndrome, is simple to operate, has high sensitivity and strong specificity, and meets the requirement of base site on rapidness and simpleness and convenience.

Description

technical field [0001] The invention belongs to the technical field of recombinant proteins, and in particular relates to a recombinant protein for detection of porcine reproductive and respiratory syndrome virus and its preparation method and application. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS), also known as "pig blue ear disease", is an acute, highly contagious infectious disease of pigs caused by porcine reproductive and respiratory syndrome virus (PRRSV). Clinically, it is mainly manifested as late abortion, premature birth, stillbirth, mummified fetus and other reproductive disorders in pregnant sows and severe respiratory diseases in pigs of various ages, especially piglets. The disease first occurred in the United States in 1987, and has since occurred in major pig-raising countries. It is now distributed worldwide, causing serious harm to the pig industry. PRRSV was isolated for the first time in my country in 1996, and it...

Claims

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Application Information

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IPC IPC(8): C07K16/46C12N15/62C12N15/13C12N15/70G01N33/569
Inventor 吴健敏覃绍敏刘金凤李作生韦建兴陈凤莲白安斌马玲饶桂波
Owner GUANGXI VETERINARY RES INST
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