欧洲型猪繁殖与呼吸综合征病毒荧光RT-PCR检测方法的建立  

Real-time RT-PCR assays for the detection of European strain of porcine reproductive and respiratory syndrome virus

作　　者：鲁韦韦[1] 唐泰山[2] 姜平[1] 张常印[2] 王凯民[2] 

机构地区：[1]南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏南京210095 [2]江苏出入境检验检疫局,江苏南京210001

出　　处：《畜牧与兽医》2008年第7期26-30,共5页Animal Husbandry & Veterinary Medicine

基　　金：江苏省科技重大项目(BE2007342);教育部博士点基金项目(20060307007);国家科技支撑计划[2007BAD86B02-3];江苏检验检疫局科研项目(No.2005KJ52)

摘　　要：根据欧洲型猪繁殖与呼吸综合征病毒(PRRSV)基因序列,设计E1和E6 2套PCR引物和TaqMan荧光探针,建立E1和E6荧光RT-PCR方法,用于检测PRRSV。反应条件优化后,用10倍倍比稀释的质粒DNA、cDNA进行PCR扩增以检测其灵敏度,同时对猪瘟(CSFV)、猪伪狂犬(PRV)等7种病毒进行特异性检测。结果显示,建立的E1和E6荧光RT-PCR可以检出欧洲型PRRSV,敏感性依次为616和216个拷贝的PRRSV重组质粒DNA,而CSFV等非PRRSV均为阴性。建立的E1、E6荧光RT-PCR具有快速、灵敏、准确、低污染等优点,可作为检测欧洲型PRRSV的技术储备。Two pairs of real-time RT-PCR primers and probes, E1 and E6, were designed and synthesized based on the nucleotide sequences of European strain of porcine reproductive and respiratory syndrome virus （PRRSV）. After the reaction conditions were optimized, El and E6 real-time RT-PCR were developed here for PRRSV. The maximal detection limits of E1 and E6 real-time RT-PCR were 616 and 216 copies of plasmid DNA containing the gene of PRRSV, respectively. The high specificity of the two methods was confirmed by detecting 7 viruses, such as classical swine fever virus （CSFV）, pseudorabies virus （PRV）, etc. The real-time RT-PCR can be used for PRRSV inspection and research.

关 键 词：猪繁殖与呼吸综合征病毒 荧光RT-PCR 检测 

分 类 号：S852.65[农业科学—基础兽医学]