猪瘟病毒荧光定量RT-PCR和RT-LAMP快速检测方法的建立  

Development of Real-time RT-PCR and RT-LAMP for CSFV Detection

作　　者：唐景峰[1] 周康平[1] 韩振伟[1] 王业富[1] 

机构地区：[1]武汉大学生命科学学院,湖北武汉430072

出　　处：《动物医学进展》2014年第3期1-5,共5页Progress In Veterinary Medicine

基　　金：质检公益性行业科研专项(201110036-03)

摘　　要：建立了应用TaqMan-LNA的荧光定量RT-PCR和反转录-环介导等温扩增(RT-LAMP)技术检测猪瘟病毒(CSFV)的两种方法。根据对NCBI中CSFV E2基因序列进行分析,设计了CSFV E2基因的引物和探针,并以E2基因为模板构建了重组质粒。通过梯度稀释阳性标准品进行试验表明荧光定量RTPCR和RT-LAMP检测下限分别为102拷贝/μL和101拷贝/μL;特异性试验表明,两种方法对于其他5种病毒不能检出,具有良好的特异性。临床样本检测同样表明两种方法都具有良好的特异性和灵敏度。说明所建立的Taq Man-LNA荧光定量RT-PCR和RT-LAMP可应用于CSFV的检测。Classical swine fever virus is one of the worldwide causative agent for the disease of pigs and causes heavy loss every year.In this study,a TaqMan-LNA based real-time RT-PCR assay and a reverse-transcription loop-mediated isothermal amplification （RT-LAMP）assay were established.The detection limits were 102 copy/μL and 101 copy/μL for the real-time PCR and LAMP assay.Two methods both showed good specificity.Futhermore,two methods were analysed with clinical samples,and they exhibited high sensitivity and specificity.The real-time RT-PCR and RT-LAMP assays will provide useful tools for the detection of CSFV.

关 键 词：猪瘟病毒 荧光定量RT-PCR 环介导等温扩增技术 

分 类 号：S852.651[农业科学—基础兽医学]