油菜花叶病毒RT-LAMP快速检测方法的建立
Establishment of RT-LAMP method for rapid detection ofyoucai mosaic virus
秦艳红 1王凤丽 1文艺 1赵正伟 2高素霞 1张德胜 1李绍建 1刘永康 2刘玉霞 1鲁书豪 1王飞 1鲁传涛1
作者信息
- 1. 河南省农业科学院植物保护研究所,郑州 450002
- 2. 温县农业科学研究所,温县 475004
- 折叠
摘要
油菜花叶病毒(youcai mosaic virus,YoMV)是侵染地黄和山药的重要病毒之一,能导致中药材的产量下降及品质降低.本研究基于逆转录环介导恒温扩增技术(reverse transcription loop-mediated isothermal amplification,RT-LAMP),建立了针对YoMV的快速、特异、灵敏的RT-LAMP检测方法.根据YoMV外壳蛋白(coat protein,CP)的核苷酸序列设计了 5条特异性引物,建立的RT-LAMP方法最适检测温度为60 ℃,反应时间为60 min,该方法能特异性检测到YoMV,灵敏度是常规PCR的1 000倍,可检测9.18×102拷贝·μL-1模板,本研究建立的RT-LAMP检测技术为地黄和山药上YoMV的准确鉴定提供了快速、高效的方法.
Abstract
Youcai mosaic virus(YoMV)is one of the important viruses infecting Rehmannia glutinosa and Dioscorea oppositifolia.It decreases the yield and quality of these Chinese medicinal herbs.Based on reverse transcription loop-mediated isothermal amplification(RT-LAMP),a rapid,specific and sensitive method for detecting YoMV was developed.Five specific primers were designed according to the nucleotide sequence of YoMV coat protein(CP).The established RT-LAMP method for detecting YoMV was optimally performed at 60 ℃ for 60 min.This method can detect YoMV specifically.The sensitivity was 1 000 times higher than that of the conventional PCR,and it could detect 9.18×102 copies·μL-1.The RT-LAMP method established in this study provided a rapid and efficient diagnostic tool for correct detection of YoMV on Rehmannia glutinosa and Dioscorea oppositifolia.
关键词
地黄/山药/油菜花叶病毒/逆转录环介导恒温扩增/快速检测Key words
Rehmannia glutinosa/Dioscorea oppositifolia/youcai mosaic virus/reverse transcription loop-mediated isothermal amplification(RT-LAMP)/rapid detection引用本文复制引用
基金项目
国家中药材产业技术体系建设项目(CARS-21)
河南省中药材产业技术体系建设项目(HARS-22-11-Z1)
河南省农科院基本科研业务费项目(2023ZC051)
出版年
2024
NETL
NSTL
万方数据