TaqMan探针荧光PCR检测肠出血性大肠埃希菌O157:H7方法的建立  

Establishment of TaqMan Probe Fluorescent PCR to Detect Enterohemorrhagic Escherichia coli O157:H7

作　　者：刘生峰[1] 肖进文[1] 刘利成[2] 杨俊[1] 王昱[1] 聂福平[1] 周庆[1] 李应国[1,3] 

机构地区：[1]重庆出入境检验检疫局,重庆400020 [2]北京华大基因研究中心,北京101300 [3]重庆市进出口食品安全工程研究中心,重庆400020

出　　处：《微生物学杂志》2012年第5期6-10,共5页Journal of Microbiology

基　　金：国家质检总局公益性项目(201010045);重庆市科技攻关项目(CSTC 2011AC1056)

摘　　要：肠出血性大肠埃希菌(Enterohemorrhagie Escherichia coli,EHEC)O157:H7是一种重要的传染病病原菌,以EHEC O157:H7标准菌株rfbE保守区设计一对特异引物和一条探针,建立了检测EHEC O157:H7核酸的荧光定量PCR检测方法。实验结果表明荧光定量PCR检测方法特异性好,最低检测限为20 cfu/mL,线性范围是102～108cfu/mL。稳定性试验表明批内变异系数和批间变异系数分别为2.06%和2.45%。Enterohemorrhagic Escherichia coli（EHEC） O157：H7 is an important infectious pathogen.A method of real-time PCR with TaqMan probe for detection of EHEC O157：H7 was developed.A pair of primers and a probe were designed based on its rfbE conservative sequences of EHEC O157：H7 standard strain,to establish fluorescent quantitative PCR（FQPCR） detection method to detect EHEC O157：H7.The results showed that the FQPCR had a good specificity with a lowest detection limit of 20 cfu/mL and linear range of 102 ~108 cfu/mL.The stability test showed that the coefficients of variation（cv） from intra-assay and inter-assay were 2.06% and 2.45% respectively.

关 键 词：肠出血性大肠埃希菌O157:H7 荧光PCR 检测 

分 类 号：Q939.93[生物学—微生物学]