畜禽肉沙门氏菌和大肠杆菌O157多重PCR检测研究

Studies on Detection of Salmonella spp. and Escherichia coli O157 in Meat and Other Animal Products by Multiplex PCR Assay

摘要:

沙门氏菌和大肠杆菌O157都是目前世界公认引起食源性疾病的重要致病菌。本研究针对致病菌传统检测方法耗时长、过程繁琐的缺点, 建立了同时检测畜禽肉及其制品中沙门氏菌和大肠杆菌O157的多重PCR分子检测方法。结果表明：分别针对沙门氏菌侵袭基因invA、大肠杆菌O157抗原基因rfbE建立的多重PCR方法可简便、快速、灵敏地实现对沙门氏菌和大肠杆菌O157的同时检测, 整个过程在9 h~10 h内完成, 人工污染猪肉检测限分别达到2.4×102 cfu/mL(沙门氏菌)和2.2×102 cfu/mL(大肠杆菌O157); 为食源性致病菌的检测提供了理想手段, 有良好的应用前景。

关键词:畜禽肉, 多重PCR, 沙门氏菌, 大肠杆菌O157

Abstract:

A rapid multiplex PCR (m-PCR) method that allows the simultaneous detection,in a single tube, of two commonly encountered food-borne pathogens in meat and other animal products was developed. The invasion protein gene (invA) of Salmonella spp. and rfbE gene of Escherichia coli O157 were used as the gene targets. The multiplex PCR assay was specific and rapid,with a turnround time of 9 h~10 h.While the detection limit is 2.4×102 cfu/mL of Salmonella spp. and 2.2×102 cfu/mL of E.coli O157 respectively when detecting the artificially contaminated pork meat with incubation at 37℃ for 4 h. The m-PCR assay developed in this study could provide a cost-effective and informative supplement to conventional microbiological methods for routine monitoring of food.

Key words:Meat, Multiplex PCR, Salmonella spp., Escherichia coli O157