I型牛疱疹病毒gE基因TaqMan探针荧光定量PCR检测方法建立  

ESTABLISHMENT OF TAQMAN PROBE REAL-TIME QUANTITATIVE PCR TO DETECT BOVINE HERPESVIRUS TYPE I GE GENE

作　　者：张晓锋[1,2] 翁永刚 陈冈[3] 李守富[3] 郭海[3] 叶结平[3] 范红结 陈鸿军[4] ZHANG Xiao-feng;WENG Yong-gang;CHEN Gang;LI Shou-fu;GUO Hai;YE Jie-ping;FAN Hong-jie;CHEN Hong-jun(Animal Medicine School,Nnajing Agricultural University,Nanjing 210095,China;Jinshan Municipal Agricultural Commission,Shanghai 201500,China;Jinshan Municipal Center for Disease Control and Prevention,Shanghai 201500,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)

机构地区：[1]南京农业大学动物医学院,南京210095 [2]上海市金山区农业委员会,上海201500 [3]上海市金山区动物疫病预防控制中心,上海201500 [4]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2018年第5期82-85,共4页Chinese Journal of Animal Infectious Diseases

摘　　要：以I型牛疱疹病毒(Bovine herpesvirus-1,BHV-1)的gE基因序列,针对1607～1704 bp区域分别设计1对引物及相应的TaqMan探针,在建立和优化反应体系后,对经10倍系列稀释的病毒进行扩增来检测其灵敏度和特异性。结果表明,建立的荧光定量PCR可用于检测BHV-1,其灵敏度为10 copies/μL,且与其他病毒无交叉反应,可用于BHV-1临床检测。To accurately establish TaqMan probe real-time quantitative PCR(qPCR)method for Bovine herpesvirus(BHV-1),the primers and TaqMan probe specific to 1607-1704 bp of gE gene was designed according to the whole genomic sequence of BHV-1 representative strain.Using the viral DNA standard template,the stability,specificity,and sensitivity of the qPCR method were investigated.The results showed that,in the standard curve,R2 value was 0.999 with a high specificity.The sensitivity of the real-time PCR was less than 10 copies/μL.No cross reactions appeared to the other herpesviruses.The TaqMan probe qPCR method has the advantages to establish the BHV-1 detection method with high sensitivity and specificity.

关 键 词：牛疱疹病毒 TaqMan实时荧光定量PCR GE基因 

分 类 号：S852.659.1[农业科学—基础兽医学]