FTA滤膜用于多重PCR检测豆制品中3种食源性致病菌的研究  

FTA Filter-based Multiplex PCR Detection of Three Species of Food-borne Pathogens in Soybean Products

作　　者：周巍 周正 康素芬 刘东 穆燕魁 

机构地区：[1]河北省食品质量监督检验研究院,河北省食品安全实验室,河北石家庄050051

出　　处：《食品科学》2009年第18期317-320,共4页Food Science

摘　　要：为建立豆制品中金黄色葡萄球菌、沙门氏菌、福氏志贺氏菌的多重PCR检测方法,采用FTA滤膜从豆制品中直接提取模板DNA,根据金黄色葡萄球菌的nuc基因、沙门氏菌的phoP基因、福氏志贺氏菌的ipaH基因,设计3对特异性引物进行多重PCR,并对反应条件进行优化。结果表明:3对引物能特异性扩增出280、409、326bp的目的条带;不增菌的情况下,多重PCR同时检测3种致病菌的灵敏度分别是101、101、102CFU/ml,检测时间4h。建立的三重PCR具有准确、快速、高效的特点,为同时检测豆制品中金黄色葡萄球菌、沙门氏菌、福氏志贺氏菌提供了新的方法。To establish a multiplex PCR method for the simultaneous detection of pathogens （Staphylococcus aureus, Salmonella spp, and Shigella flexneri） in soybean products, FTA filter was used to directly extract the template DNAs from soybean products. On the basis of nuc gene of Staphylococcus aureus, phoP gene of Salmonella spp, ipaH gene of Shigella flexneri, three pairs of specific primers were designed for the multiplex PCR detection, and the reaction conditions were optimized. The results showed that three targeted genes of 280, 409, and 326 bp could be specifically amplified using the three pairs of primers.Without bacterial enrichment, the sensitivities of the multiplex PCR for detecting Staphylococcus aureus, Salmonella spp, and Shigella flexneri were 10^1, 10^1, and 10^2 CFU/ml, respectively, and the detection could be finished within 4 h. This multiplex PCR method is accurate, fast and effective, thereby providing a new approach for the detection of Staphylococcus aureus, Salmonella spp, and Shigellaflexneri in soybean products.

关 键 词：多重PCR 食源性致病菌 检测 FTA滤膜 

分 类 号：TS201.3[轻工技术与工程—食品科学]