双启动寡核苷酸聚合酶链式反应法检测食品中的沙门氏菌  

Detection of Salmonella in food by dual priming oligonucleotide-polymerase chain reaction

作　　者：郭华麟 徐超 李轲 胡雪慧 李飞雨 钟婷婷 韩国全[1] 

机构地区：[1]四川农业大学食品学院,雅安625014 [2]河南出入境检验检疫局检验检疫技术中心,郑州450003

出　　处：《食品安全质量检测学报》2017年第10期4004-4008,共5页Journal of Food Safety and Quality

基　　金：成都市生鲜猪肉病原体携带风险监测研究项目(CD2015004);2016年度四川农业大学本科生科研兴趣培养计划项目(2016182)~~

摘　　要：目的建立检测食品中沙门氏菌的双启动寡核苷酸-聚合酶链式反应(dual priming oligonucleotide-polymerase chain reaction,DPO-PCR)方法。方法以沙门氏菌invA基因为靶基因设计一对DPO引物,优化条件,对180份熟肉样本同时进行DPO-PCR和细菌分离鉴定。结果 DPO-PCR方法退火温度在52、57、62℃均可对靶基因高效扩增。方法特异性好,仅对沙门氏菌为阳性结果,而对金黄色葡萄球菌、志贺氏菌、大肠杆菌O157:H7、β溶血性链球菌、铜绿假单孢菌无交叉反应。方法灵敏度可达4.3×10~2 CFU/mL。与细菌分离鉴定相比,两者检测结果完全一致。结论该方法快速、精准,可作为食品中沙门氏菌的快速检测方法。Objective To establish a dual priming oligonucleotide-polymerase chain reaction（DPO-PCR） method for determination of Salmonella in food. Methods Based on the invA gene of Salmonella, a pair of DPO primer was designed. The reaction conditions were optimized. A total of 180 cooked meat samples were detected by bacteria isolation and identification and DPO-PCR method in parallel. Results The results showed that the target gene was efficiently amplified by DPO-PCR at annealing temperatures of 52, 57 and 62 ℃. The method was specific, only the results of Salmonella were positive, and no cross reactions with Staphylococcus aureus, Shigella, Escherichia coli O157：H7, β Hemolytic streptococcus, and Pseudomonas aeruginosa. The sensitivity of the method was 4.3×10^2 CFU/mL. Compared with bacterial isolation and identification, the test results were exactly the same. Conclusion This method is rapid and accurate, which can be used as a rapid detection method of salmonella in food.

关 键 词：沙门氏菌 invA基因 双启动寡核苷酸-聚合酶链式反应 

分 类 号：TS207.4[轻工技术与工程—食品科学]