牛轮状病毒TaqMan实时荧光RT-PCR快速检测方法的建立  

Detection of Bovine Rotavirus by TaqMan Based Real-time Reverse Transcription Polymerase Chain Reaction Assay

作　　者：范晴[1] 谢芝勋[1] 刘加波[1] 庞耀珊[1] 邓显文[1] 谢志勤[1] 谢丽基[1] 彭宜[1] 

机构地区：[1]广西兽医研究所,广西南宁530001

出　　处：《中国畜牧兽医》2011年第4期105-108,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家百千万人才工程人选专项资金资助(945200603);广西科技攻关与新产品试制项目(桂科合0992033-5)

摘　　要：本研究按照牛轮状病毒（BRV）结构蛋白VP6基因序列,设计合成引物和探针,经各反应条件的优化,建立了BRVTaqMan实时荧光定量RT-PCR技术。对BRV进行了特异性、敏感性和重复性试验。结果表明,TaqMan实时荧光RT-PCR最低可检测到100个拷贝病毒RNA;与牛病毒性腹泻病毒（BVD）、猪瘟病毒（CSFV）、牛结核杆菌（MB）和牛传染性鼻气管炎病毒（IBRV）不发生交叉反应;所制作的标准曲线在102~109拷贝/μL浓度范围内有极好的线性关系且线性范围宽,相关系数为0.997;与常规的RT-PCR相比,该方法具有快速、特异、敏感、重复性好、可同时检测大量样品等优点。可对样品中微量BRV进行准确检测,对BRV的诊断有重要意义。A TaqMan based real-time RT-PCR was developed for the specific detection of bovine rotavirus（BRV）.The specific primers and probes were designed according to the VP6 sequences of BRV.It was found that the specificity of this assay was high without any cross-reactions and had no CSFV,BVD,MB and IBRV.A good linear correlation was demonstrated in the standard curve for the real-time PCR assay within the range of 102 to 109copies,with a correlation coefficient of 0.997.The detection limit of the real-time RT-PCR assay was 100 copies of viral RNA,indicating a good sensitivity of the assay.This TaqMan based real-time RT-PCR assay reported here is a valuable method with high specificity and sensitivity detection of BRV.

关 键 词：牛轮状病毒 TaqMan实时荧光定量PCR 检测 

分 类 号：Q78[生物学—分子生物学]