多重PCR检测食品中的金黄色葡萄球菌、志贺菌和沙门菌  

Detection of Staphylococcus aureus, Shigella spp.,Salmonella spp.in food by multiplex PCR

作　　者：李博[1] 陈福生[1] 王小红[1] 邵彦春[1] 朱胜梅[1] 

机构地区：[1]华中农业大学食品科技学院,武汉430070

出　　处：《卫生研究》2008年第4期438-442,共5页Journal of Hygiene Research

基　　金：华中农业大学"大学生科技创新基金"(SRF)重点资助项目

摘　　要：目的建立一种能同时检测食品中金黄色葡萄球菌、志贺菌和沙门菌的多重PCR检测方法。方法采用7.5%NaCl肉汤对食品样品中的金黄色葡萄球菌进行增菌,同时采用GN增菌剂对食品样品中的志贺菌和沙门菌进行增菌。根据金黄色葡萄球菌的nuc基因、志贺菌的ipaH基因、沙门菌的invA基因设计引物,通过多重聚合酶链反应(PCR)反应对食品样品中上述三种病原菌的目标基因进行扩增,同时对反应体系进行优化。结果特异性实验表明本方法的特异性良好。对污染金黄色葡萄球菌、志贺菌和沙门菌的牛奶样品进行检测,检出限为1cfu/ml。结论本实验建立的多重PCR检测方法适用于食品中金黄色葡萄球菌、志贺菌和沙门菌的快速检测,具有快速、简便、灵敏的特点。Objective To establish a multiplex PCR method for simultaneous detection of Staphylococcus aureus, Shigella spp,, Salmonella spp, in food, Methods Staphylococcus aureus was enriched by 7.5 % NaCl broth while Shigella spp, and Salmonella spp. were enriched by GN medium , The primers were designed according to the gene nuc of Staphylococcus aureus, the gene ipaH of Shigella spp. and the gene invA of Salmonella spp,, The target genes of these pathogens in food were amplified by multiplex PCR, which reaction conditions were optimized specifically, Results The multiplex PCR method established in this experment was of high specificity, which detection limit was 1 cfu /ml of Staphylococcus aureus, Shigella spp, and Salmonella spp, when the milk samples contaminated with these pathogens. Conclusion The multiplex PCR method, which was rapid, convenient, and with high sensitivity, could be suitable for rapid detection of Staphylococcus aureus, Shigella spp,, Salmonella spp, in food , and could have a great prospect.

关 键 词：金黄色葡萄球菌 志贺菌 沙门菌 多重聚合酶链反应 食品检测 

分 类 号：R155.51[医药卫生—营养与食品卫生学]