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

中文关键词:  沙门氏菌  invA基因  双启动寡核苷酸-聚合酶链式反应

英文关键词:Salmonella  invA gene  dual priming oligonucleotide-polymerase chain reaction

中文摘要:

      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4.3×102 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。

英文摘要:

      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 4.3×102 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.