羊传染性脓疱病毒半套式PCR检测方法的建立及应用  

Development and Application of Semi-nested PCR for Orf Virus Detection

作　　者：邓宇[1] 

机构地区：[1]西昌学院动物科学学院,四川西昌615013

出　　处：《动物医学进展》2014年第8期28-31,共4页Progress In Veterinary Medicine

基　　金：西昌学院博士科研启动基金项目(13BQZ05);西昌学院校县横向合作项目(XA0807)

摘　　要：根据羊传染性脓疱病毒F1L基因序列设计3条引物,建立了一种能鉴别诊断羊传染性脓疱病毒和绵羊痘病毒、山羊痘病毒的半套式PCR快速检测方法。采用该方法对羊传染脓疱病毒XC13株进行检测,结果半套式PCR扩增出450bp的特异性DNA片段,而对山羊痘病毒、绵羊痘病毒、禽痘病毒的扩增结果均为阴性;敏感性试验结果表明,所建立的PCR方法能检出3.64×101 copies/μL羊传染性脓疱病毒;重复性试验、临床样品检测试验结果表明,所建立的半套式PCR检测方法可用于羊传染性脓疱病毒感染的快速诊断与流行病学调查。According to the sequence of F1 Lgene of Orf virus in GenBank,three primers were designed and synthesized.A semi-nested PCR assay for differentiating Orf virus（ORFV）,sheep pox virus（SPV）and goat pox virus（GPV）was developed.The 450 bp DNA fragment was amplified in ORFV XC13 strain,but no bands were amplified with templates extracted respectively from SPV,GPV and avian pox virus.Sensitivity of the semi-nested PCR assay was 3.64×10^1 copies/μL ORFV.In the repeatability test and the field trail,the semi-nested PCR assay could be used as a method for the detection and molecular epidemiological investigation of Orf virus.

关 键 词：半套式PCR 羊传染性脓疱病毒 绵羊痘病毒 山羊痘病毒 

分 类 号：S852.659.1[农业科学—基础兽医学] Q789[农业科学—兽医学]