基于HAND系统5种致腹泻大肠杆菌多重实时PCR初筛方法的建立  

Establishment of multiplex real-time PCR assays based on HAND system for the preliminary screening of five diarrheagenic Escherichia coli

作　　者：钟宜科 王永霞 赵彤 贺晓明 郭建树 刘威 邹大阳 ZHONG Yi-ke;WANG Yong-xia;ZHAO Tong;HE Xiao-ming;GUO Jianshu;LIU Wei;ZOU Da-yang(Hebei University of Engineering,College of Life Sciences and Food Engineering,Handan,Hebei 056038,China;Chinese PLA Center for Disease Control and Prevention,Beijing 100000,China)

机构地区：[1]河北工程大学生命科学与食品工程学院,河北邯郸056038 [2]中国人民解放军疾病预防控制中心,北京100000

出　　处：《食品与机械》2019年第10期46-51,共6页Food and Machinery

基　　金：国家科技重大专项（编号：2018ZX10713-003）;河北省邯郸市科技攻关项目（编号：1722201063-3)

摘　　要：以实验室17株种属关系较近的菌株进行特异性评价,通过溶解曲线T m值对扩增产物进行分析表明,该方法具有高特异性,可同时检出5种致泻性大肠杆菌EAEC(aggR)、EHEC/EPEC(eae)、ETEC(LT)和EIEC(ipaH)的相关毒力基因。粪便模拟样本试验表明,其敏感性达10^4~10^6 CFU/mL。建立的多重实时PCR检测方法适用于5种致泻性大肠杆菌的初步筛选。The specificity of this method was evaluated by testing 17 species closely related bacteria in the laboratory,and the amplification product was analyzed by the Tm value of the dissolution curve,which had good specificity and could detect five different virulence genes:aggR for EAEC,eae for EHEC and EPEC,LT for ETEC,ipaH for EIEC in the same reaction tube.The fecal simulated samples showed that the sensitivity reached 10 ^4~10^6 CFU/mL.The multiplex real-time PCR detection method established in this experiment is suitable for the preliminary screening of five kinds of Escherichia coli.

关 键 词：致泻性大肠杆菌 多重实时PCR 一管化 

分 类 号：S85[农业科学—兽医学]