PRRSV和PCV-2以及PRV多重SYBR Green-Ⅰ实时荧光PCR检测方法的建立  

Establishment of a multiplex SYBR Green-Ⅰ real time PCR assay for simultaneous detection of porcine reproductive and respiratory syndrome virus,porcine circovirus type 2 and porcine pseudorabies virus

作　　者：王小武[1] 符芳[1] 柴政[1] 孔令达[1] 蔡雪辉[1] 宋淑萍[1] 许红喜[2] 李曦[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室 猪传染病研究室,黑龙江哈尔滨150001 [2]黑龙江八一农垦大学,黑龙江大庆163319

出　　处：《中国兽医科学》2008年第6期494-499,共6页Chinese Veterinary Science

基　　金：黑龙江省科技攻关计划重点项目(GB05B501-1)

摘　　要：根据GenBank中登录的猪生殖与呼吸综合征病毒(PRRSV)N蛋白基因、猪圆环病毒2型(PCV-2)rep蛋白基因和猪伪狂犬病病毒(PRV)gE基因的核苷酸序列分别设计了3对特异性引物,成功建立了同时检测PRRSV、PCV-2、PRV的多重SYBR Green-Ⅰ实时荧光PCR方法。敏感性试验结果显示,PRRSV、PCV-2的敏感性可达250拷贝/μL,PRV的敏感性可达500拷贝/μL。表明,该方法具有较好的特异性、重复性和敏感性,可以用于PRRSV、PCV-2和PRV的快速检测。Three pairs of primers were designed respectively based on the sequences of N protein gene of PRRSV, rep protein gene of PCV-2 and gE gene of PRV available in GenBank. A multiplex SYBR Green-Ⅰ real time PCR assay was established successfully for simultaneous detection of the 3 viruses. The sensitivities for PRRSV,PCV-2 and PRV were 250,250 and 500 copies/μL, respectively. The results revealed that the established PCR assay was sensitive, specific and reproducible and it could be used to detect rapidly PRRSV,PCV-2 and PRV.

关 键 词：猪生殖与呼吸综合征病毒 猪圆环病毒2型 猪伪狂犬病病毒 多重SYBR Green-Ⅰ实时荧光 PCR 

分 类 号：S852.659.6[农业科学—基础兽医学]