双重PCR方法检测沙门氏菌和空肠弯曲菌

Development of a Duplex PCR for Detection of Salmonella spp and Campylobacter jejuni in Food

为建立能够同时检测食品中沙门氏菌和空肠弯曲菌的双重PCR方法。采用沙门氏菌鞭毛基因fimY和空肠弯曲菌马尿酸酶基因hipO设计特异性引物,并对影响PCR扩增的主要因素——引物浓度、退火温度、Mg2+浓度因素进行优化,比较单一PCR和双重PCR的检测效果。结果表明:采用单一PCR法检测沙门氏菌和空肠弯曲菌时,灵敏度分别可达到3.98pg和4.05pg;而采用双重PCR检测时,灵敏度较单一PCR法有所下降,沙门氏菌和空肠弯曲菌检出限量分别为398pg和40.5pg。本研究建立的特异性强和灵敏度高的双重PCR检测方法,可为实现食品中沙门氏菌和空肠弯曲菌的同时检测提供新方法。

A rapid assay for simultaneous detection of Salmonella spp and Campylobacter jejuni in food was developed. The primers were designed based on the fimY gene of Salmonella spp and hipO gene of Campylobacter jejuni. Annealing temperatures, Mg2＋ concentrations, and primer concentrations were optimized. The detection limit was 3.98 pg for Salmonella spp and 4.05 pg for Campylobacter jejuni using the single PCR method. However, the detection limit of the duplex PCR method was significantly higher, which was 398 pg, 40.5 pg for Salmonella spp and Campylobacter jejuni, respectively. In this study, a duplex PCR with high specificity and sensitivity was developed and it would provide useful information for the simultaneous detection of Salmonella spp and Campylobacter jejuni in food.