羊传染性脓疱病毒SYBR,Green,I实时荧光定量PCR检测方法的建立
摘要：根据GenBank公布的羊传染性脓疱病毒VIR基因序列 （No. KF6665631），利用Beacon Designer 79软件设计1对特异性引物，建立羊传染性脓疱病毒VIR基因SYBR Green I实时荧光定量PCR检测方法。结果显示，该方法可以特异性检测羊传染性脓疱病毒，对羊常见病原无特异性扩增；最低检测限为100 copies>μL-1；组内和组间变异系数均小于2%；应用该方法对39份临床样品进行检测，阳性率为949%（37/39）。以上结果表明该方法特异性强、灵敏度高、重复性好，适合于羊传染性脓疱的病原学检测和流行病学调查。
关键词：羊传染性脓疱病毒；SYBR Green I；实时荧光定量PCR
中图分类号：S 85826文献标识码：A文章编号：1008-0384（2018）04-341-05
Development and Application of a Realtime Fluorescence Quantitative PCR for Detection of Orf Virus
LIN Yusheng，JIANG Jinxiu，ZHANG Jingpeng，JIANG Bin，YOU Wei，HU Qilin*
（Institute of Animal Husbandry & Veterinary Medicine，Fujian Academy of Agricultural Sciences，Fuzhou，Fujian350013，China）
Abstract：The study established a SYBR Green I realtime fluorescence quantitative PCR assay for detecting orf virus. Using Beacon Designer 79 software to design a pair of specific primers based on the VIR gene of the orf virus in the Genbank （accession number.KF6665631）. The result showed that it would specifically detect orf virus，but wouldn"t detect other sheep and goat bacteria and viruses. The detection limit of the method was 100 copies>μL-1 and the detected Ct values of intraand intervariation were all less than 2%. Using the assay to detect 39 clinical samples，the positive rate was 949% （37/39）. The above results indicated that the assay had a high specificity，sensitivity and repeatability，which could be used to detect orf virus and conduct epidemiological investigation.
Key words：orf virus；SYBR Green I；realtime fluorescence quantitative PCR
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