Brucellosis rapid detection method
A technology for rapid determination of brucellosis, applied in the field of determination, can solve problems such as low success rate, harsh conditions for isolation and identification of pathogens, cumbersome CFT operation, etc., shorten detection time, reduce operation steps and labor intensity, and overcome detection The long-term effect
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[0011] The rapid assay method for brucellosis of the present invention comprises the following steps: 1) expression fragment cloning: boil the S2 live bacteria vaccine in boiling water for 30 minutes, and then take 1 μL-2 μL as a template for PCR amplification of the target gene expression fragment, Use primers LFGCGAACCGGCAATACCAG and LBGCCGCGTTGAGTACCGT as front and rear primer pairs to carry out PCR reaction, use DNA gel recovery kit to purify and recover the target fragment, and recover the lysate of Brucella Omp19 outer membrane protein gene expression fragment; 2) Escherichia coli prokaryotic expression vector The construction of the method: take the recovered Brucella Omp19 outer membrane protein gene expression fragment lysate and the Escherichia coli pET-28a plasmid vector lysate in step (1) and carry out double enzyme digestion treatment with endonuclease EcoRI and XhoI respectively, and then , T4 DNA ligase ligation in a metal bath at 16°C for 12 hours to obtain the ...
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