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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。
The purpose is to establish a multiplex PCR method that can simultaneously detect Staphylococcus aureus, Salmonella and Shigella. Methods Three pairs of primers specific to the nuc gene of Staphylococcus aureus, invA of Salmonella and ipaH gene of Shigella were designed and used to obtain 223 bp, 302 bp and 369 bp amplified fragments by PCR. The amplified products were confirmed by sequencing For the purpose of fragment. Results The established FTA filter combined with multiplex PCR detection method simultaneously detected these three kinds of foodborne pathogens, the sensitivity reached 102 cfu / ml. Conclusion The method has high sensitivity and short time-consuming and can be used for the simultaneous detection of three kinds of pathogenic bacteria. It provides a new detection method for the prevention and control of the outbreak of bacterial food poisoning.