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Hog cholera indirect hemagglutination detection kit and preparation

A detection kit and swine fever technology, which is applied in the field of recombinant protein preparation of swine fever antigen, can solve the problems of not being able to distinguish BVDV infection, and achieve the effects of saving time, practical cost, reducing obstacles, and good stability

Inactive Publication Date: 2009-07-22
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Claims
  • Application Information

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Problems solved by technology

Currently, conventional immunological methods using CSF whole virus antigens to detect serum antibodies are unable to differentiate BVDV infection except for the use of monoclonal antibodies against CSFV and BVDV respectively to distinguish them

Method used

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  • Hog cholera indirect hemagglutination detection kit and preparation
  • Hog cholera indirect hemagglutination detection kit and preparation

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Embodiment Construction

[0020] First, compare the CSFV and BVDV sequences published on the Genbank database with DNAstar software, and select the most conserved 60 nucleotide sequences in CSFV. The principle of selecting nucleotides is: the selected nucleotide sequence must be the most conservative in CSFV; it must include at least one antigenic site that can stimulate the animal body to produce neutralizing antibodies, so as to ensure that the recombinant protein expressed later can be compatible with The anti-CSFV serum reacts; the homology of the amino acid encoded by the sequence of the selected nucleotide and the amino acid encoded by the BVDV nucleotide sequence of the corresponding region is less than 35%, so as to ensure that the recombinant protein expressed subsequently has no anti-BVDV serum Cross-reactivity; the codons in the selected nucleotide sequence should avoid the occurrence of rare codons or sub-rare codons as much as possible to ensure the smooth expression of recombinant proteins...

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Abstract

The invention relates to a preparation method of a recombinant protein, an indirect classical swine fever hemagglutination test kit that is prepared by the protein, and a preparation method of the test kit. The preparation method of the protein comprises following steps: a segment with the gene homeology degree comparing with a bovine viral diarrhea-mucosal disease virus (BVDV) that is lower than 35 percent is selected from a conservative region of a classical swine fever virus (CSFV) gene, thus obtaining a nucleotide sequence S1 that is corresponding to an amino acid A1; a forth amino acid in the nucleotide sequence S1 is modified to an hydrophilic amino acid, thus obtaining an amino acid A2; a corresponding nucleotide sequence S2 of A2 is modified, thus respectively obtaining three new nucleotide sequences; the three new nucleotide sequences are orderly and serially connected by connecting nucleotides, thus obtaining a sequence S9 that is corresponding to an amino acid sequence A3; and a nucleotide sequence S10 is obtained by processing S9 correspondingly, and after artificially synthesizing and digesting S10, connecting S10 with a carrier pGEX-4T-1 and inducing the recombinant carrier into colon bacillus for induction expression, the recombinant protein is obtained by separating and purifying an expression product.

Description

technical field [0001] The invention relates to a method for preparing a recombinant protein of hog fever antigen and a method for preparing an indirect hemagglutination detection kit by using the recombinant protein of hog fever antigen and the kit. Background technique [0002] Swine fever is a viral disease that seriously threatens the swine industry and causes significant economic losses. It is listed as one of the 16 legal infectious diseases of Class A by the World Organization for Animal Health (OIE). Announcement No. 96 issued by the Ministry of Agriculture of my country on February 12, 1999 also listed swine fever as a first-class animal disease. [0003] So far, swine fever has existed for nearly 2 centuries. Although countries all over the world have made great efforts to prevent and control the disease, and successfully eliminated swine fever in some areas of North America, Europe and Northern Europe, swine fever It is still widely spread in many parts of the wo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C07K14/18G01N33/86C12R1/19
Inventor 郑福英邱昌庆蔺国珍周继章曹小安宫晓炜
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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