| 鲁书豪1, 秦艳红1, 杨会珍2, 杨 瑾1, 李雪梦1, 李绍建1, 文 艺1, 高素霞1, 王 飞1*, 鲁传涛1*.地黄葱X病毒RT-LAMP快速检测方法的建立与应用[J].植物保护,2026,(1):266-272. |
| 地黄葱X病毒RT-LAMP快速检测方法的建立与应用 |
| Establishment and application of RT-LAMP rapid detection method for Rehmannia allexivirus |
| 投稿时间：2025-01-21 修订日期：2025-03-31 |
| DOI：10.16688/j.zwbh.2025041 |
| 中文关键词: 地黄 地黄葱X病毒 反转录环介导等温扩增(RT-LAMP) 可视化 快速检测 |
| 英文关键词:Rehmannia glutinosa Rehmannia allexivirus RT-LAMP visualization fast detection |
| 基金项目:国家中药材产业技术体系（CARS-B21）；河南省中药材产业技术体系（HARS-22-11-Z1）；河南省农业科学院基本科研业务费（2024ZC051） |
| 作者 | 单位 | E-mail | | 鲁书豪1, 秦艳红1, 杨会珍2, 杨 瑾1, 李雪梦1, 李绍建1, 文 艺1, 高素霞1, 王 飞1*, 鲁传涛1* | 1. 河南省农业科学院植物保护研究所, 郑州 450002
2. 方城县植保植检站, 南阳 473200 | 王飞yunfeiren@163.com；鲁传涛chuantaolu@qq.com |
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| 中文摘要: |
| 地黄葱X病毒(Rehmannia allexivirus, ReAV)是首次在地黄上报道的新病毒, 本研究基于ReAV外壳蛋白(coat protein, CP)基因的核苷酸序列设计了3组引物进行筛选, 对RT-LAMP进行了温度优化、时间优化、特异性检测、灵敏度比较等试验, 建立了ReAV反转录环介导恒温扩增(reverse transcription loop-mediated isothermal amplification, RT-LAMP)快速检测技术。结果表明:本研究建立的ReAV RT-LAMP方法最适反应条件为65℃ 40 min, 该方法可特异性检测出ReAV，其检测限为6.99拷贝/μL, 灵敏度是常规PCR的10倍。对21份地黄样品的检测结果表明, RT-LAMP方法的阳性检出率(85.7%)略高于常规RT-PCR方法(71.4%)。本研究建立的RT-LAMP检测技术为ReAV的准确检测提供了快速、特异、简便的方法, 为田间样品的检测提供了技术支撑。 |
| 英文摘要: |
| Rehmannia allexivirus (ReAV) is a new virus that firstly reported on Rehmannia glutinosa. In this study, three pairs of primers were designed based on nucleotide sequence of coat protein (CP) gene of ReAV for primer screening. Temperature optimization, time optimization, specificity detection and sensitivity comparison of RT-LAMP were carried out. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid detection of ReAV was established. The results showed that the optimal conditions for the RT-LAMP method were 65℃ for 40 min. The optimized LAMP method could specifically detect ReAV, and there was no cross reaction with other viruses on R.glutinosa. The sensitivity of RT-LAMP was 10 times higher than that of conventional PCR, and 6.99 copies/μL template could be detected. In 21 R.glutinosa samples, the RT-LAMP assay demonstrated higher sensitivity (85.7%) than conventional RT-PCR (71.4%) for detecting ReAV. The RT-LAMP method developed in this study enables rapid, specific and simple detection of ReAV, providing a practical tool for field detection. |
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