食品中3种致病菌多重PCR检测方法的建立及初步应用  

Development and application of a multiplex PCR assay for detection of three pathogenic bacteria in food

作　　者：周蓓莉[1,2] 肖进文[2,3] 刘生峰[2,3] 李应国[2,3] 周庆[2] 聂福平[2] 王昱[2] 杨俊[2] 

机构地区：[1]西南大学食品科学学院,重庆400715 [2]重庆出入境检验检疫局,重庆400202 [3]重庆市进出口食品安全工程技术研究中心,重庆400202

出　　处：《食品科技》2012年第6期312-315,共4页Food Science and Technology

基　　金：重庆市科技攻关项目(CSTC;2011AC1056)

摘　　要：建立用多重聚合酶链式反应(Multiplex polymerase chain reaction,mPCR)同时检测食品中沙门氏菌、金黄色葡萄球菌和单核细胞增生性李斯特菌的方法。以沙门氏菌的gyrB基因、单核细胞增生性李斯特菌的gyrB基因、金黄色葡萄球菌的coa基因作为目的基因,分别设计3对引物,通过优化反应体系,建立3种致病菌的多重PCR检测体系。采用单重PCR检测时,灵敏度可达0.423ng/mL(沙门氏菌)、2.5ng/mL(金黄色葡萄球菌)和0.36ng/mL(单核细胞增生性李斯特菌);而采用三重PCR检测时,灵敏度较单重PCR有所下降,分别为2.43ng/mL(沙门氏菌)、2.5ng/mL(金黄色葡萄球菌)、3.6ng/mL(单核细胞增生性李斯特菌)。初步建立能同时、快速、灵敏地检测食品中沙门氏菌、金黄色葡萄球菌和单核细胞增生性李斯特菌的三重PCR方法。A multiplex polymerase chain reaction（mPCR） assay for the simultaneous detection of Salmonella spp., Staphylococcus aureus and Listeria monocytogenes in food was developed in this study. Three pairs of primers were designed respectively according to the gryB gene of Salmonella spp., the coa gene of Staphylococcus aureus, the gryB gene of Listeria monocytogen. Multiplex PCR was established by optimizing the reaction system. Results： The detection limit was 0.423 ng/mL for Salmonella spp., 2.5 ng/mL for Staphylococcus aureus, 0.36 ng/mL for Listeria monocytogenes using the single PCR method. However, the sensitivity of the multiple PCR method was higher, which was 2.43 ng/mL for Salmonella spp, 2.5 ng/mL for Staphylococcus aureus, 3.6 ng/mL for Listeria monocytogenes respectively. A triplePCR assay has been established for the simultaneous, rapid and sensitive detection of Salmonella spp., Staphylococcus aureus and Listeria monocytogenes in food.

关 键 词：食源性致病菌 多重聚合酶链式反应(mPCR) 食品 检测 

分 类 号：TS207.4[轻工技术与工程—食品科学]