三种食源性致病菌的多重PCR 快速检测及应用.PDF

生物技术通报 ·技术与方法· BIOTECHNOLOGY BULLETIN 年第 期 2014 7 三种食源性致病菌的多重 PCR 快速检测及应用 余倩  黄梦娜 （仲恺农业工程学院 轻工食品学院，广州 510225） 摘 要 ： 针对常见的3 种食源性致病菌肠出血性大肠杆菌O157 H7 的 基因、金黄色葡萄球菌的 基因及沙门氏菌 rfbE nuc 的hilA 基因，使用Primer 5.0 软件设计出相对应的特异性引物，预计PCR 产物的分子大小为287 、354 及468 bp 。通过单一、双重 及三重PCR 对样品进行检测，并对人工感染的鲜奶进行检测。结果表明，3 种食源性致病菌的单一、双重和三重PCR 均能成功扩 增出与预计大小一致的片段，无非特异性扩增。人工感染食品的PCR 检测也获得了目的菌的特异性片段，并且结果稳定。这说明 此3 对引物可分别用于3 个菌靶基因的PCR 检测。 关键词 ： 食源性致病菌 肠出血性大肠杆菌O157 H7 金黄色葡萄球菌 沙门氏菌 PCR - Detection and Application of Three Food borne Bacterial Pathogens by Multiplex PCR Yu Qian Huang Mengna （Zhongkai University of Agiculture and Engineering ，Guangzhou 510225 ） Abstract:  Food poisoning incidents have occurred by the pollution of food-borne pathogenic bacteria frequently. PCR meet the requirements of the rapid detection of pathogenic bacteria. The three primers was designed by primer 5.0 software of Enterohemorrhagic O157 H7 gene, gene, gene. Three amplified segments of gene, gene, Escherichia coli rfbE Staphyloccocus aureus nuc Salmonella hilA rfbE nuc hilA gene were 287 bp, 354 bp, 468 bp. The specificity and sensitivity of primers were verified by single, duplex and triplex-PCR reaction, and by detecting the foods artificially infected with the three pathogens. The results sh