利用多重PCR检测3种食源性致病菌

Detection of three food-borne bacterial pathogens by using multiplex PCR

目的旨在建立一种可以同时检测金黄色葡萄球菌、沙门氏菌和志贺氏菌的多重PCR方法。方法以金黄色葡萄球菌的nuc基因、沙门氏菌invA、志贺氏菌ipaH基因作为靶序列设计3对特异性引物,进行PCR反应得到223bp、302 bp、369 bp扩增片段,经测序证实扩增产物为目的片段。结果采用建立的FTA滤膜结合多重PCR的检测方法同时对这3种食源性致病菌进行检测,灵敏度均达到102 cfu/ml。结论该方法灵敏度高、耗时短,可用于同时检测3种致病菌,为预防控制细菌性食物中毒的暴发流行提供了新的检测方法。

Objective To establish a multiplex PCR Shigella spp and Salmonella spp at the same time. for the simultaneous detection of Staphylococcus aureus, Methods Primer targeting the Staphylococcus aureus nuc gene, Salmonella invA gene, Shigella spp ipaH gene were used in the multiplex PCR, three DNA fragments of 223bp, 302bp and 369bp were amplified, confirmed by sequencing of amplified products for the purpose. Re- suits FTA filter was used to extract pathogens DNA, the three pathogens were simultaneously detected by the multiplex PCR technology which has been designed, the sensitivity of the multiplex PCR was 102 cfu/mL for the three pathogens. Conclusions The multiplex PCR method lies on its accuracy, rapidity and efficiency in the diagnosis, so it could be a useful method for the simultaneous detection of the three species of bacteria in food and the prevention of bacterial food poisoning.