首页|油菜花叶病毒RT-LAMP快速检测方法的建立

油菜花叶病毒RT-LAMP快速检测方法的建立

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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.
Establishment of RT-LAMP method for rapid detection ofyoucai mosaic virus
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.

Rehmannia glutinosaDioscorea oppositifoliayoucai mosaic virusreverse transcription loop-mediated isothermal amplification(RT-LAMP)rapid detection

秦艳红、王凤丽、文艺、赵正伟、高素霞、张德胜、李绍建、刘永康、刘玉霞、鲁书豪、王飞、鲁传涛

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河南省农业科学院植物保护研究所,郑州 450002

温县农业科学研究所,温县 475004

地黄 山药 油菜花叶病毒 逆转录环介导恒温扩增 快速检测

国家中药材产业技术体系建设项目河南省中药材产业技术体系建设项目河南省农科院基本科研业务费项目

CARS-21HARS-22-11-Z12023ZC051

2024

10.13926/j.cnki.apps.001336

植物病理学报
中国植物病理学会

植物病理学报

CSTPCD北大核心
影响因子：0.865
ISSN：0412-0914
年,卷(期)：2024.54(4)
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