Double-antibody sandwich ELISA kit for detecting porcine acute diarrhea syndrome coronavirus
A technology for acute diarrhea and coronavirus, applied in the field of double-antibody sandwich ELISA kits, can solve the problems of unsuitability for rapid detection, expensive equipment, long test cycle, etc., and achieve rapid detection methods and monitoring methods with good specificity
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[0056] In the following, the present invention will be described in more detail through specific embodiments, so as to facilitate the understanding of the technical solution of the present invention, but it is not used to limit the protection scope of the present invention.
[0057] The SADS-CoV GDS04 strain was donated by Professor Cao Yongchang of Sun Yat-sen University.
[0058] 1. Construction, expression and purification of SADS-CoVN protein recombinant plasmid
[0059] 1.1 Construction of pET32a-N recombinant plasmid
[0060] Using the N protein of SADS-CoV GDS04 strain (Genbank accession number: MF167434.1) as a reference, design specific primers, upstream P1: 5′-CGC GGATCC ATGGCCACTGTTAATTGG-3′, downstream P2: 5′-CCG CTCGAG CTAATTAATAATCTCATC-3', BamHI and XhoI restriction sites (underlined part) were introduced at the 5' ends of the upstream and downstream primers. After the primers were synthesized, PCR amplification was performed using the SADS-CoV GDS04 strain...
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