基于RT-RAA和CRISPR/Cas12a的一管式玉米矮花叶病毒快速检测方法  

A one-tube rapid detection method for maize dwarf mosaic virus based on RT-RAA and CRISPR/Cas12a

作　　者：侯雨萌 赵振兴 王思元 董铮 胡中泽[3] 周涛 张永江[1] HOU Yumeng;ZHAO Zhenxing;WANG Siyuan;DONG Zheng;HU Zhongze;ZHOU Tao;ZHANG Yongjiang(Chinese Academy of Inspection and Quarantine,Beijing 100176,China;College of Plant Protection,China Agricultural University,Beijing 100193,China;Taizhou Institute of Agricultural Sciences,Jiangsu Academy of Agricultural Sciences,Taizhou 225300,China)

机构地区：[1]中国检验检疫科学研究院,北京100176 [2]中国农业大学植物保护学院,北京100193 [3]江苏省农业科学院泰州农业科学研究所,江苏泰州225300

出　　处：《江苏农业学报》2025年第2期268-275,共8页Jiangsu Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2021YFD1400100、2021YFD1400103)。

摘　　要：玉米矮花叶病毒(Maize dwarf mosaic virus,MDMV)是一种重要的检疫性病毒,严重影响玉米的生产。为了有效防治玉米矮花叶病,本研究基于反转录重组酶介导等温核酸扩增(RT-RAA)技术和CRISPR/Cas12a系统构建了一种MDMV快速检测方法,将所有试剂集中在一个试管中进行反应,并通过侧向流动试纸条(LFD)检测反应结果。本研究筛选得到的最佳反应条件为,引物添加量2.8μL、FB报告分子浓度100 nmol/L、RT-RAA系统反应时间20 min、CRISPR/Cas12a系统反应时间20 min。本研究构建的基于RT-RAA和CRISPR/Cas12a的一管式玉米矮花叶病毒快速检测方法特异性强、灵敏度高,且所用试验材料方便携带和运输,适用于现场检测。Maize dwarf mosaic virus(MDMV)is an important quarantine virus that seriously affects maize production.To effectively prevent and control maize dwarf mosaic disease,this study constructed a rapid detection method for MDMV based on reverse transcription-recombinase aided amplification(RT-RAA)technology and the CRISPR/Cas12a system.All reagents were reacted in a single test tube,and the reaction results were detected by lateral flow dipstick(LFD).The optimal reaction conditions screened in this study were as follows:primer addition amount of 2.8μL,FB reporter molecule concentration of 100 mol/L,RT-RAA system reaction time of 20 minutes,and CRISPR/Cas12a system reaction time of 20 minutes.The one-tube rapid detection method for maize dwarf mosaic virus based on RT-RAA and CRISPR/Cas12a constructed in this study has strong specificity,high sensitivity,and the experimental materials used are convenient for carrying and transportation,making it suitable for on-site detection.

关 键 词：玉米矮花叶病毒 反转录重组酶介导等温核酸扩增(RT-RAA)技术 CRISPR/Cas12a 侧向流动试纸条 

分 类 号：S513[农业科学—作物学]