TaqMan探针荧光RT-PCR检测狂犬病病毒  

Detection of Rabies Virus by TaqMan Probe Real-time RT-PCR

作　　者：王振全[1,2] 罗宝正[2] 薄清如[2] 周昌芳[1] 白鸽[1] 许远靖[1] 王冲[1] 吴殿君[1] 

机构地区：[1]吉林大学农学部畜牧兽医学院,吉林长春130062 [2]珠海出入境检验检疫局国家外来病检测重点实验室,广东珠海519015

出　　处：《中国畜牧兽医》2012年第6期79-82,共4页China Animal Husbandry & Veterinary Medicine

基　　金：广东省科技计划项目(2009B030801075)

摘　　要：根据GenBank已公布的狂犬病病毒(rabies virus,RV)核蛋白(N)基因序列设计并合成一对特异性的引物和探针,建立基于TaqMan探针的荧光RT-PCR检测狂犬病病毒方法。对狂犬病疫苗提取核酸后进行RT-PCR扩增,将目的条带切胶回收,克隆测序,重组质粒作为标准阳性对照。对建立的TaqMan探针荧光RT-PCR方法做灵敏度、特异性、重复性及稳定性试验。结果显示,该方法可以达到10拷贝/μL的灵敏度,可以将狂犬病病毒与犬瘟热病毒、犬细小病毒、犬腺病毒、犬冠状病毒和犬副流感病毒分开,方法重复性好,稳定可靠。To establish the method of TaqMan probe real-time RT-PCR for detection of rabies virus, a pair of primers and probe were designed based on the nucleoprotein sequences of rabies virus that published in GenBank. RNA was extracted from rabies virus vaccine, and then amplified by RT-PCR, cloned the target fragment and took the recombination plasmid as stand- ard positive control. The results showed that the method can distinguished rabies virus from canine distemper virus, canine parvovirus, canine adenoviruses, canine coronavirus and canine para influenza virus, the sensitivity could attained 10 copies/μL and the stability test was good.

关 键 词：狂犬病病毒 TAQMAN探针 荧光RT-PCR 

分 类 号：S854.4[农业科学—临床兽医学]