关键词: 多重PCR, 沙门氏菌肺, 炎克雷伯杆菌, 嗜肺巴斯德菌, 铜绿假单胞菌

Abstract: Objective To establish a protocol for simultaneous detection of four common pathogens in SPF environment. Method A multiplex PCR assay was established with primers designed according to invA gene of Salmonella typhimurium, khe gene of Klebsiella pneumoniae, 16S rRNA gene of pasteurella pneumotropica and ecfX gene of Pseudomonas aeruginosa. The specificity and sensitivity of the new multiplex PCR system were also evaluated after optimization. Result The multiplex PCR method could simultaneously amplify the four bacteria in the same sample without cross reaction, and the minimum detection concentration was 10-3ng/μL. The multiplex PCR method could also specifically detect the target pathogens from co-infection disease material. Conclusion The multiplex PCR method , which was rapid, convenient and sensitivity, could provide effective technical for the rapid diagnosis and monitoring of these four pathogens.

Key words: multiplex PCR, Salmonella typhimurium, Klebsiella pneumoniae, pasteurella pneumotropica, Pseudomonas aeruginosa