5种食源性致病菌多重PCR检测方法的建立  

Multiplex PCR for the detection of five major foodborne pathogens

作　　者：程晓艳[1,2] 刘庆慧[1] 黄倢 

机构地区：[1]农业部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所,青岛266071 [2]中国海洋大学,青岛266003

出　　处：《渔业科学进展》2012年第2期97-103,共7页Progress in Fishery Sciences

基　　金：国家自然科学基金(30871942);对虾行业专项(201103034)课题共同资助

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病原菌副溶血弧菌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 parahaemolyticus,Staphylococcus aureus,Listeria monocytogenes,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. monocytogenes, 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.

关 键 词：多重PCR 食源性病原菌 检测 

分 类 号：S917.1[农业科学—水产科学]