Porcine circovirus disease detection method based on ORF2 (Open Reading Frame 2) antigen domain
A technology of GST-ORF2-E and functional area, which is applied in botany equipment and methods, biochemical equipment and methods, viral peptides, etc., can solve the problems of time-consuming and labor-intensive, virus-dispersing infected cells, etc., to improve sensitivity and specificity , the effect of improving specificity
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[0029] 1. Primer design and PCR amplification of target gene
[0030] Data report (MahéD, Blanchard P, Truong C, Arnauld C, Le Cann P, CarioletR, Madec F, Albina E, Jestin A. 2000. Differential recognition of ORF2protein from type 1 and type 2porcine circoviruses and identification of immunorelevant epitopes. J Gen Virol. 81 (Pt 7): 1815-1824.), there are five main immune response regions on the protein encoded by PCV2, one of which is on the ORF1 protein coding region, and the other four on ORF2. Because only the antigen functional area (113-147) on ORF2 is the most effective, it may become the main antigen for the development of ELISA kits for detecting PCV2 infection in herds. In the present invention, the amino acid segment (113-147) on ORF2 is selected as the target antigen for cloning and expression, and its gene fragment is 102 bp in length. Using the whole PCV2 ORF2 gene as a template, two specific primers were designed, and the gene fragments located in the 113-147aa sp...
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