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.
作者
张晓锋
翁永刚
陈冈
李守富
郭海
叶结平
范红结
陈鸿军
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)
出处
《中国动物传染病学报》
CAS
北大核心
2018年第5期82-85,共4页
Chinese Journal of Animal Infectious Diseases
作者简介
张晓锋,男,高级兽医师,博士,主要从事农产品质量安全、农业科技推广和重大动物疫病防控应急管理;通信作者:陈鸿军,E-mail:vetchj@shvri.ac.cn
