美洲型猪繁殖与呼吸综合征病毒TaqMan荧光RT-PCR检测方法的建立  

Establishment of TaqMan real-time RT-PCR assays for detection of north american type porcine reproductive and respiratory syndrome virus

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

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：《中国预防兽医学报》2008年第5期389-392,共4页Chinese Journal of Preventive 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);国家自然科学基金(30471288);国家科技支撑计划(2006BAD06A04);江苏检验检疫局科研项目(2005KJ52)

摘　　要：根据猪繁殖与呼吸综合征病毒(PRRSV)美洲株的基因序列,设计U1和U6两套PCR引物和荧光探针,建立了扩增U1和U6基因的荧光RT-PCR用于检测美洲型PRRSV。用10倍倍比稀释的质粒DNA、cDNA、RNA进行扩增以检测其灵敏度,同时对猪瘟(HCV)、猪伪狂犬(PRV)等7种病毒进行特异性检测。结果显示建立的扩增U1和U6基因的荧光RT-PCR可用于检测PRRSV,其灵敏度为10个拷贝的质粒DNA,而与HCV等非PRRSV无交叉反应。本研究所建立的荧光RT-PCR具有快速、灵敏、准确、低污染等优点,可用于PRRSV样品的检测。Two pairs of real-time RT-PCR primers and probes, U1 and U6 were designed and synthesized based on the nucleotide sequences of the porcine reproductive and respiratory syndrome virus （PRRSV）. The minimal detection threshold of U1 and U6 real-time RT-PCR were approximately 10 copies of plasmid DNA, The specificity of the two methods was demonstrated by detecting 7 viruses including classical swine fever virus （HCV）, pseudorabies virus （PRV）. It was concluded that the real time RT-PCR could be effective methods for rapid detection of PRRSV.

关 键 词：猪繁殖与呼吸综合征病毒 荧光RT-PCR 特异性 灵敏度 

分 类 号：S852.65[农业科学—基础兽医学]