猪戊型肝炎病毒实时荧光定量PCR检测方法的建立及初步应用  

Establishment and application of a real-time fluorescence PCR assay for detecting swine hepatitis E virus

作　　者：邱蜜蜜[1] 孟轲音[2] 丁壮[1] 叶俊青[1] 丛彦龙[1] 李志杰[1] 刘美[1] 李少丽[1] 王昌庆[1] 吴昊[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2010年第4期449-452,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30771606)

摘　　要：根据GenBank中注册的AJ272108等序列,参考有关文献,利用引物设计相关软件在HEV ORF2片段的保守区设计并合成1对引物及探针,采用TaqMan探针建立了荧光定量PCR法。以鉴定正确的质粒为模板建立标准曲线,Ct值线性范围为14.89~27.02,相关系数为0.996。本试验建立的猪戊型肝炎病毒ORF2片段基因实时荧光定量PCR方法扩增效率高、线性范围广,为快速检测猪戊型肝炎病毒的绝对定量分析奠定了基础。According to registered AJ272108 sequence and other related sequences in GenBank,and referring the literatures of Meng and other researchers,a pair of primers and the probe were designed and synthesized for the conservative region of HEV ORF2 fragment,and a real-time fluorescence PCR assay was developed by the TaqMan probe.The standard curve was established though using the positive plasmid as template,in which the linear range of Ct value was from 14.89 to 27.02,and correlation coefficient was 0.996.The real-time fluorescence quantitative PCR assay with the efficient amplification and the wide linear range laid a foundation for rapidly detecting the absolute quantitative analysis of swine hepatitis E virus.

关 键 词：猪戊型肝炎 实时荧光定量PCR TAQMAN探针 

分 类 号：S852.65[农业科学—基础兽医学]