2009•Jiangsu nongye xuebaoRequires access
Method of multiplex-PCR for identification of Pasteurella multocida and Haemophilus parasuis.
He Ying, Ping Zhao, Yuefeng Chu, Pengcheng Gao, Jianjun Zhang, LU Zhong-xin
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Abstract
Multiplex-PCR detection method for Pasteurella multocida(PM) and Haemophilus parasuis(HPS) had been developed based on published single PCR for these pathogens.The PCR reagents,annealing temperature and the PCR procedure were optimized and the primers were screened,the PCR products including 457 bp of PM and 1 090 bp of HPS were obtained just by one test.The detectable minimum dose of HPS was 1.04×10~3 CFU or 7.2 pg DNA and PM was 2.3×10~3 CFU or 16 pg DNA.Among 23 suspected isolates,two strains showed PM fragments by multiplex-PCR method, no HPS fragments,but all artificially infected isolates showed HPS fragments.The results suggested that the multiplex-PCR method could be used in clinical diagnosis of PM and HPS.