5种食源性致病菌多重PCR检测方法的建立

摘要

本研究建立一种同时特异性地检测5种常见食源性致病菌的多重PCR方法.根据目前食品中常见痛原菌副溶血弧菌Vibrio parahaemolyticus、金黄色葡萄球菌Staphylococcus aureus、单 核细胞增生性李斯特菌Listeria monocytogenes、肠炎沙门氏菌Salmonella enteritidis、福氏志贺氏菌Shigella flexneri的相关毒力基因,选择具有特异性的副溶血弧菌不耐热溶血毒素基因、金黄色葡萄球菌的耐热核酸酶基因、单核细胞增生性李斯特菌编码溶血素O基因、沙门氏菌侵袭蛋白A基因及志贺氏菌侵袭性质粒抗原H基因,设计5对特异性引物进行多重PCR检测,对反应条件进行优化并评价了其特异性和灵敏度.通过对48份实际样品检测,验证了此多重PCR体系具有很好的可靠性和实用性.%A multiplex polymerase chain reaction was developed for the simultaneous detection of five major foodborne pathogens,Vibrio parakaemolyticus,Staphylococcus aureus,Listeria monocy to genes,Salmonella enteritidi,and Shigella flexneri.Five pairs of specific primers were designed according to the thermolabile hemolysin gene of V.parahaemolyticus,the heat stable nuclease gene of S.aureus,the Listeriolysin O gene of L.monocy to genes,the invasion protein A gene of S.enteritidis,the invasion plasmid antigen H gene of S.flexneri.The reaction conditions were optimized,and the specificity and sensitivity of this method were tested.The detecting results of 48 samples also proved its reliability and practicability.