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Kit and detection method for detecting titer of European PRRSV antibody

An antibody titer, European-type technology, applied in the field of molecular biology, can solve the problem of the gap between natural active proteins and other problems, and achieve the effect of high-efficiency soluble expression and good immunoreactivity

Inactive Publication Date: 2020-08-07
SOUTH CHINA AGRI UNIV
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Due to the relatively simple operation and low cost of preparation, Escherichia coli has become the preferred system for exogenous gene expression in vitro, but most of the expressed exogenous proteins exist in the form of insoluble inclusion bodies, which are far from natural active proteins, which limits its use. application

Method used

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  • Kit and detection method for detecting titer of European PRRSV antibody
  • Kit and detection method for detecting titer of European PRRSV antibody
  • Kit and detection method for detecting titer of European PRRSV antibody

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Embodiment Construction

[0079] In order to make the objectives, technical solutions and advantages of the present invention clearer, the following is attached Figure 1-6 And examples, the present invention will be further described in detail. It should be understood that the specific embodiments described herein are only used to explain the present invention, but not to limit the present invention.

[0080] 1. Material

[0081] (1) PCR amplification template and serum

[0082] Plasmid pMD-18T-PRRSV-ORF7 containing European PRRSV-ORF7, anti-European PRRSV N protein rabbit serum, European and American PRRSV positive pig serum and PRRSV negative pig serum, porcine circovirus type 2 (PCV2) positive Pig serum, classical swine fever virus (CSFV) positive pig serum, pseudorabies virus (PRV) positive pig serum, porcine parvovirus (PPV) positive pig serum.

[0083] (2) Reagent

[0084] PET-32a carrier, anti-His mouse monoclonal antibody and Factor Xa were purchased from NEB; HRP-labeled anti-mouse IgG antibody, HRP-...

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Abstract

The invention provides a nucleic acid molecule for encoding a PRRSV N antigen and an application thereof, and further provides a kit for detecting the titer of an European PRRSV antibody and a detection method. The detection kit provided by the invention is high in practicability, capable of realizing high-throughput operation, capable of saving the detection cost and convenient to use. Clinical detection results show that the indirect ELISA detection method established by taking the N protein as the coating antigen is strong in specificity, and lays a foundation for continuously researching the European porcine reproductive and respiratory syndrome virus.

Description

Technical field [0001] The invention relates to the technical field of molecular biology, in particular to a nucleic acid molecule encoding PRRSV N antigen and its application, a kit for detecting the titer of a European PRRSV antibody, and a detection method. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRS virus, PRRSV), which can cause reproductive disorders in sows and fattening pigs Respiratory diseases, accompanied by mild neurological symptoms, can cause systemic viremia in severe cases. In recent years, due to genetic recombination, new strains (such as NADC30, NADC30-like) have emerged, making the prevention and control of PRRS more difficult. According to the genetic characteristics of the virus and the global epidemic situation, PRRSV can be divided into type 1 (European type) and type 2 (American type). Type 2 is mainly preva...

Claims

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Application Information

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IPC IPC(8): C12N15/40C12N15/62C07K14/08C07K19/00C12N15/70G01N33/68G01N33/569G01N33/543
CPCC07K14/005C07K2319/21C12N15/70C12N2770/10022C12N2770/10051G01N33/54306G01N33/56983G01N33/6854G01N2333/08G01N2469/20
Inventor 贺东生蔡蔚游陈湘杨心怡司广斌周霞李锦辉李根冯军森韦柳明梁文清陈志飞陈一波
Owner SOUTH CHINA AGRI UNIV
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