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African swine fever virus antigen recombinant tandem epitope protein, construction method and application thereof, and antibody ELISA kit

An African swine fever virus and a construction method technology, applied in the field of antibody ELISA kits, can solve problems such as reducing the replication ability of ASFVBA71 virus strain, and achieve the effects of good repeatability and stability, wide diagnostic range and high sensitivity

Pending Publication Date: 2022-07-22
HUNAN AGRICULTURAL UNIV +1
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Problems solved by technology

[0007] ASFV MGF genes (such as MGF360 and MGF505, etc.) encode multigene family (MGF) proteins, and it has been reported that MGF360 and MGF505 genes are involved in inhibiting the production of interferon (IFN); the deletion of MGF genes can reduce the expression of ASFV BA71 virus strain in porcine macrophages. replication capability in

Method used

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  • African swine fever virus antigen recombinant tandem epitope protein, construction method and application thereof, and antibody ELISA kit
  • African swine fever virus antigen recombinant tandem epitope protein, construction method and application thereof, and antibody ELISA kit
  • African swine fever virus antigen recombinant tandem epitope protein, construction method and application thereof, and antibody ELISA kit

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Embodiment

[0045] An African swine fever virus antigen recombinant tandem epitope protein, which is formed by connecting the African swine fever virus CD2v protein and the B cell epitope of the MGF360-505R protein with a flexible peptide (ie MGF360-505R-CD2v), and its amino acid sequence is as SEQID As shown in NO: 1, the nucleotide sequence encoding the MGF360-505R-CD2v is shown in SEQ ID NO: 2.

[0046] The construction and screening process of the African swine fever virus antigen recombinant tandem epitope protein is as follows (such as figure 1 shown):

[0047] 1. Selection (collection or prediction) of p30, p72, CD2v and MGF360-505R epitopes of African swine fever virus (ASFV epidemic strain in China)

[0048] Download p30, p72 and CD2v of ASFV Chinese prevalent strains (SY18, HLJ, DB / LN, wbBS01, Anhui, CAS19, Wuhan1 and Wuhan2) from NCBI (https: / / www.ncbi.nlm.nih.gov / ), respectively Sequences, the GenBank accession numbers of ASFV circulating strains in China are: SY18(MH766894)...

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Abstract

The invention discloses an African swine fever virus antigen recombinant tandem epitope protein, which is formed by connecting B cell epitopes of an African swine fever virus CD2v protein and an MGF360-505R protein in series through a flexible peptide, and the tandem epitope protein has higher expression quantity and inclusion body content, has certain solubility, has reactivity with a serum antibody, and can be used for antibody ELISA (Enzyme-Linked Immunosorbent Assay) detection. The invention also discloses a construction method of the African swine fever virus antigen recombinant tandem epitope protein, an application of the African swine fever virus antigen recombinant tandem epitope protein in antibody ELISA detection and an antibody ELISA detection kit. According to the application and the antibody ELISA detection kit, the specificity, repeatability and stability are good, the sensitivity is high, serum samples generated after ASFV wild type and CD2v gene deletion type strain immunization can be accurately recognized, a certain reference value is achieved for ASF detection, and a serological detection method based on conservative epitopes is expected to provide a wider diagnosis range.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to an African swine fever virus antigen recombinant tandem epitope protein MGF360-505R-CD2v and its construction method, application and antibody ELISA kit. Background technique [0002] African swine fever (ASF) is a highly lethal swine infectious disease caused by African swine fever virus (ASFV). The particle structure of African swine fever virus is similar to that of iridescent virus, and the replication mode has the same characteristics as poxviruses. It is the only member of the African swine fever virus family. According to the classification report, the African swine fever virus family has been classified into the order Asfuvirales and Pokkesviricetes. [0003] In-depth research on the serodiagnostic targets of ASF will help to explore its infection detection, pathogenic mechanism and immune system response. The high conservation of ASFV important...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N15/66G01N33/68G01N33/569G01N33/58G01N33/543
CPCC07K14/005C12N15/70G01N33/6854G01N33/56983G01N33/581G01N33/54393C12N2710/12022C12N2710/12051C07K2319/40G01N2333/01G01N2469/20
Inventor 王乃东杨文兵邹亚文张丽杰邬静杨青王昌建金业
Owner HUNAN AGRICULTURAL UNIV
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