关键词: 山羊地方性鼻内肿瘤病毒； TaqMan； 荧光定量RT?PCR

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