猪瘟病毒TaqMan荧光定量PCR检测方法的建立  

Development of TaqMan fluorescent quantitative PCR method for detecting classical swine fever virus(CSFV)

作　　者：马琳[1] 付薇[1] 何奇松[2] 徐贤坤[1] 易春华[2] 孙翔翔[2] 蒋家霞[2] 黄胜斌[1] 熊毅[1] 

机构地区：[1]广西动物疫病预防控制中心,南宁530001 [2]广西大学动物科学技术学院,南宁530005

出　　处：《广西农业科学》2010年第11期1223-1225,共3页Guangxi Agricultural Sciences

基　　金：广西科技攻关项目(桂科攻0815009-3-7)

摘　　要：根据GenBank中5′端非编码区的靶序列设计了用于TaqMan荧光定量PCR的1对引物及TaqMan探针,以猪瘟活疫苗病毒为模板,在常规PCR的基础上优化了TaqMan荧光定量PCR反应条件,建立了猪瘟病毒(CSFV)TaqMan荧光定量PCR检测方法。其敏感性和特异性试验结果表明,所建立的TaqMan荧光定量PCR检测方法对CSFV具有很高的特异性,其检测灵敏度可达5.03×10-2μg/μL。说明TaqMan荧光定量PCR适用于CSFV的快速检测。A pair of primers and a TaqMan probe were designed according to the CSFV 5′UTR sequences of CSFV taken from GenBank,for performing TaqMan fluorescent quantitative PCR.RNA of classical swine fever virus（CSFV） was used as template,and the reaction conditions of TaqMan fluorescence PCR were optimized based on conventional PCR.The TaqMan fluorescent quantitative PCR method for CSFV was developed,which showed high specificity and high sensitivity to CSFV（detection sensitivity of 5.03×10-2 μg/μL）,as revealed by the sensitivity and specificity tests.Therefore,this method can be used for rapid detection of CSFV.

关 键 词：猪瘟病毒 TaqMan荧光定量PCR TAQMAN探针 

分 类 号：S858.28[农业科学—临床兽医学]