基于RAA技术快速检测EHEC-O157型核酸方法的建立  

Establishment of a rapid detection method for Enterohemorrhagic E.coli O157 based on fluorescence probe RAA technology

作　　者：任宝红 郭瑞 韩小改 段蕾 贾继民[3] REN Baohong;GUO Rui;HANG Xiaogai;DUAN Lei;JIA Jimin(State Key Laboratory of Market Regulation(Food Safety Rapid Testing and Smart Supervision Technology),Henan Zhengzhou 450003,China;Zhengzhou Zhongdao Biotechnology Co.,Ltd.,Henan Zhengzhou 450001,China;Center for Disease Control and Prevention of Xinjiang Military Region Security Department,Xinjiang Urumqi 830011,China)

机构地区：[1]国家市场监管重点实验室(食品安全快速检测与智慧监管技术),河南郑州450003 [2]郑州中道生物技术有限公司,河南郑州450001 [3]新疆军区保障部疾病预防控制中心,新疆乌鲁木齐830011

出　　处：《食品工程》2024年第2期72-75,共4页Food Engineering

基　　金：国家市场监管重点实验室(食品安全快速检测与智慧监管技术)科研项目(ZDSYS202306)。

摘　　要：针对肠出血性大肠埃希氏菌O157型建立一种核酸诊断方法。针对该病原的rfbE基因,利用重组酶介导等温核酸扩增(recombinase aided isothermal amplification,RAA)技术建立检测方法,并做性能评价。该方法在20 min内完成扩增,与多种标准菌株无交叉反应,质粒10 copies/μL或菌液102 CFU/mL可以检出阳性,37℃加压48 h后数据稳定。该方法在特异性、敏感性、稳定性表现良好,可用于肠出血性大肠埃希氏菌O157型检测。Establish a diagnostic method for Enterohemorrhagic E.coli O157.The detection method of rfbE gene was established by Recombinase Aided Isothermal Amplification(RAA)and its performance was evaluated.The method can complete the amplification detection within 20 min,and has no cross reaction with 5 kinds standard strains.The sensitivity of plasmid is 10 copies/μL,and nucleic acid is 102 CFU/mL.The data were stable after 48 hours of 37℃pressure.The method has good specificity,high sensitivity,good stability and fast detection speed,and can be used to detect with Enterohemorrhagic E.coli O157.

关 键 词：重组酶介导等温核酸扩增 O157 rfbE 

分 类 号：TS207.3[轻工技术与工程—食品科学]