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

中文关键词:  多重PCR  沙门氏菌  弗氏枸橼酸杆菌  奇异变形杆菌  迟缓爱德华菌

英文关键词:multiplex PCR  Salmonella spp  Citrobacter freundii  Proteus mirabilis  Edwardsiella tarda

中文摘要:

      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。

英文摘要:

      Objective To establish a method for detection of four species of food-borne pathogens synchronously by multiplex PCR. Methods Four pairs of suitable primers were designed according to the specific gene fimA of Salmonella spp, gene ldh of Citrobacter freundii , gene idsC of Proteus mirabilis, gene fimA of Edwardsiella tarda. The reaction conditions of multiplex PCR were optimized as well. Results The detec-tion 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.