摘要
根据GenBank公布的羊传染性脓疱病毒VIR基因序列(No.KF666563.1),利用Beacon Designer 7.9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100copies·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94.9%(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。
The study established a SYBR Green Ⅰ real-time fluorescence quantitative PCR assay for detecting orf virus.Using Beacon Designer 7.9 software to design a pair of specific primers based on the VIRgene of the orf virus in the Genbank（accession number.KF666563.1）.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 intra-and inter-variation were all less than 2%.Using the assay to detect 39 clinical samples,the positive rate was 94.9%（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.
作者
林裕胜
江锦秀
张靖鹏
江斌
游伟
胡奇林
LIN Yu sheng;JIANG Jin xiu;ZHANG Jing peng;JIANG Bin;YOU Wei;HU Qi lin(Institute of Animal Husbandry ＆ Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, China)
出处
《福建农业学报》
CAS
北大核心
2018年第4期341-345,共5页
Fujian Journal of Agricultural Sciences
基金
国家重点研发计划项目(2016YFD0500906)
福建省科技计划项目--省属公益类科研院所基本科研专项(2018R1023-9)
福建省财政专项--福建省农业科学院青年创新团队(STIT2017-3-10)
福建省农业科学院科技创新项目(PC2018-6)
