关键词: 地黄灼烧病毒, 地黄, 环介导等温扩增技术, 可视化检测

Abstract: Rehmannia torradovirus virus（ReTV）is a new virus first reported on Rehmannia glutinosa，which seriously endangers the growth of R.glutinosa in the field. In this study，four sets of specific primers were designed with ReTV coat protein gene as the target gene，and the RT‑LAMP detection technology of ReTV was established. Primer screening and RT‑LAMP product identification were performed using electrophoresis detection and visualization methods（with SYBR Green Ⅰ reagent）.At the same time，the RT‑LAMP reaction conditions were optimized by a single variable method.The ReTV recombinant plasmid was used as a template for specific detection and sensitivity comparison，and the field rehmannia samples were detected.The results showed that the optimal reaction condition for the established ReTV RT‑LAMP detection method was 65° C for 60 min.The specific detection results showed that only ReTV was positive as the template，and there was no cross reaction with other viruses on R.glutinosa. The sensitivity of RT‑LAMP was 100 times higher than that of conventional RT‑PCR.RT‑LAMP could detect 3.39×100 copies/μL.RT‑LAMP method had a 100% positive detection rate，and RT‑PCR method had a 93.75% positive detection rate，indicating that the accuracy of RT‑LAMP was better than that of RT‑PCR method. The RT‑LAMP detection technology established in this study provides a rapid，specific and simple method for the accurate detection of ReTV.

Key words: Rehmannia torradovirus virus, Rehmannia glutinosa, LAMP, Visual detection