多重PCR技术同时检测四种肠道致病菌方法的建立与初步应用

Establishment and primary application of multiplex polymerase chain reaction for synchronous detection of four food-borne pathogens

目的建立同时检测4种食源性肠道致病菌的多重PCR方法。方法分别依据沙门氏菌(Salmonella spp)的fimA基因、弗氏枸橼酸杆菌(Citrobacter freundii)的ldh基因、奇异变形杆菌(Proteus mirabilis)的idsC基因、迟缓爱德华菌(Edwardsiella tarda)的fimA基因设计4对特异性引物,并对多重PCR反应条件进行优化。结果建立的多重PCR方法具有结果可靠、灵敏度高、方便快捷的优点。结论本研究为研发快速检测以上4种食源性肠道致病菌的试剂盒提供了重要技术保障。

Objective To establish a method for detection of four species of food-borne pathogens syn-chronously by multiplex PCR. Methods Four pairs of suitable primers were designed according to the spe-cific gene fimA of Salmonella spp, gene ldh of Citrobacter freundii , gene idsC of Proteus mirabilis, and gene fimA of Edwardsiella tarda. The reaction conditions of multiplex PCR were optimized as well. Results The detection for real samples proved that the multiplex PCR method had the advantages of reliability, sensitivity and rapidness. Conclusion This method can provide a key technical support for developing a kit that detects the four species of food-borne pathogens rapidly.