Abstract: In order to establish a rapid detection method for Enzootic nasal tumor virus of goats（ENTV ? 2）， primers and TaqMan probes were designed according to the conserved region of ENTV?2 env gene. By optimizing the reaction conditions，the ENTV ?2 TaqMan fluorescence quantitative RT ?PCR detection method was established，and its specificity，sensitivity，repeatability and clinical detection effect were verified. The results showed that the standard curve established by this method had a correlation coefficient（R2 ）of 0.998 when the concentration of recombinant plasmid was 1.019×109 —1.019×102 copies/μL，and the Ct value had a good linear relationship with the concentration of recombinant plasmid. This approach was specific for ENTV ? 2，and there was no cross ? reaction observed against other five kinds of pathogens related to goats. The minimum detection limit was 10.19 copies/μL，andthe coefficient of variation intra ?assays and inter ?assays was within 0.46% and 1.5%. It was more sensitive than the conventional RT?PCR and gave higher ENTV?2 positive detection rate（60%，12/20）than the conventional RT?PCR （30% ，6/20）from clinical samples，the total coincidence rate of the two methods was 70%（14/20）.These data indicated that the TaqMan ? based realtime RT ? PCR assay established here was an effective method with high sensitivity，specificity and reproducibility for faster and more accurate detection and quantification of ENTV?2.

Key words: Enzootic nasal tumor virus?2； TaqMan； Real?time quantitative RT?PCR

摘要： 为建立山羊地方性鼻内肿瘤病毒（Enzootic nasal tumor virus of goats，ENTV?2）快速检 测方法，根据 ENTV?2 env 基因保守区域设计引物和 TaqMan 探针，通过优化反应条件，建立 TaqMan 荧光定量 RT?PCR 检测方法，并对其特异性、灵敏性、重复性与临床检测效果进行验 证。结果显示，该方法建立的标准曲线在重组质粒浓度为 1.019×109 —1.019×102 copies/μL时， 相关系数R2为0.998，Ct值和重组质粒浓度有良好的线性关系；与相关的其他5种羊常见病原 无交叉反应，最低检测限度为 10.19 copies/μL，组间和组内的变异系数均在1.5%内。利用本研 究建立的方法与常规RT?PCR对20份临床样品进行对比检测，两种方法的ENTV?2阳性检出率 分别为60%（12/20）和30%（6/20），两种方法的总符合率为70%（14/20）。结果表明，本研究建 立的方法特异性强、灵敏度高、重复性好，为ENTV?2的诊断和流行病学调查提供技术手段。

关键词: 山羊地方性鼻内肿瘤病毒； TaqMan； 荧光定量RT?PCR