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

摘要： 地黄灼烧病毒（Rehmannia torradovirus virus，ReTV）是首次在地黄上报道的新病毒，严重危害地黄田间生长。为建立快速检测ReTV 的反转录环介导等温扩增技术（RT-LAMP），以ReTV 外壳蛋白（Coat protein，CP）基因为靶标基因设计了4组特异性引物，利用电泳检测及可视化方法（加SYBR Green Ⅰ显色剂）进行引物筛选和RT-LAMP产物鉴定，同时以单一变量法优化RT-LAMP反应条件，以ReTV 重组质粒为模板检测特异性和灵敏度，并对田间地黄样品进行检测应用。结果表明，建立的ReTV RTLAMP 检测方法最适反应条件为65 ℃ 60 min；特异性检测结果表明，只有ReTV为模板显示阳性，与地黄上其他病毒无交叉反应；RT-LAMP灵敏度是常规RT-PCR的100倍，可检测到3.39×100拷贝/μL；在样品检测中RT-LAMP方法的阳性检出率（100%）高于常规RT-PCR方法（93.75%），表明RT-LAMP方法准确性高于常规RT-PCR方法。建立的RT-LAMP检测技术为ReTV的准确检测提供了快速、特异、简便的方法。

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