检测猪戊型肝炎病毒的荧光定量PCR方法的建立  

Establishment of a real-time fluorescent quantitative PCR assay for the detection of swine hepatitis E virus

作　　者：张亮权[1] 欧阳昀[1] 刘志刚[1] 曹宗喜[1] 闫明菲[1] 王文华[1] 郭泗虎[1] 张桂红[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医杂志》2011年第10期16-18,共3页Chinese Journal of Veterinary Medicine

基　　金：国家生猪现代农业产业技术体系(NYCYTX009);广州市科技支撑计划项目"猪戊型肝炎诊断关键技术的研究"(10A82101592)

摘　　要：根据GenBank中猪戊型肝炎病毒的ORF2核苷酸序列的保守区域设计合成一对特异性引物,建立了一套SYBRGreenⅠ荧光定量PCR检测猪源戊型肝炎病毒(swHEV)的方法,并评价了该方法的灵敏度、稳定性和特异性,同时与常规的RT-nPCR进行对比分析。结果表明,建立标准曲线的相关系数为0.998,斜率为-3.039,Ct值变异系数(CV)在0.17%～1.41%之间,有良好的稳定性。同时在检测猪群常见病中显示出很好的特异性,并且比RT-nPCR更灵敏,适合于swHEV的检测。According to the published sequences of swHEV in GenBank, a pair of primers were de- signed and synthesized for the conserved open reading frame 2 （ORF2） of HEV, and a SYBR Green I fluorescent real-time quantitative PCR assay was developed. The sensitivity, stability and specificity were evaluated, and comparison of the Real-time PCR with nested RT-PCR was performed. According to the experiment, the correlation coefficient（re）and the slope value of the standard curve was 0. 998 and --3.039 respectively. The coefficient of variation （CV） of intra-assay was between 0. 17% and 1. 41%, which showed good stability. The sensitivity and specificity of real-time PCR was higher than nested RT-PCR, which can detect swHEV efficiently.

关 键 词：猪戊型肝炎病毒 荧光定量PCR RT-nPCR 

分 类 号：S852.659.6[农业科学—基础兽医学]