葡萄扇叶病毒RT-LAMP检测方法的建立  

Establishment of RT-LAMP assay for detection of grapevinee fanleaf virus

作　　者：谈静惠 谢婷 张博文 陈林 Tan Jinghui;Xie Ting;Zhang Bowen;Chen Lin(Yinchuan Customs District,Yinchuan 750001,China)

机构地区：[1]银川海关,宁夏银川750001

出　　处：《植物检疫》2023年第4期8-12,共5页Plant Quarantine

基　　金：海关总署科研项目(2021HK177)。

摘　　要：葡萄扇叶病毒(grapevine fanleaf virus,GFLV)是侵染葡萄的主要病毒之一,严重影响葡萄的产量和品质。该病毒的传统检测方法操作复杂、耗时长,本研究利用反转录环介导等温扩增(reverse transcription loop-mediated isothermal amplification,RT-LAMP)技术,建立葡萄扇叶病毒的RT-LAMP荧光快速检测方法。研究结果显示,新建的GFLV RT-LAMP检测方法最适检测温度为64℃,在60 min内可完成反应,灵敏度为普通RT-PCR方法的10倍,且特异性良好。用GFLV的RT-LAMP方法对葡萄基地采集的90份葡萄叶片样品进行检测,结果与普通RT-PCR方法检出一致。说明建立的葡萄扇叶病毒RT-LAMP检测方法有快速、便捷、特异性强、灵敏度高的特点,可供海关检疫、植保站、实验室及其他机构使用。Grapevine fanleaf virus(GFLV)is one of the main viruses infecting grape,which seriously affects the yield and quality of grape.The traditional detection method of the GFLV is complicated and time-consuming.In this study,a RT-LAMP fluorescence rapid detection method for GFLV was established based on reverse transcription loop-mediated isothermal amplification.The results showed that the optimized RT-LAMP can be done at 64℃within 60 min.Sensitivity of the RT-LAMP was 10 times higher than that of traditional RT-PCR and the specificity was good.The eastablished RT-LAMP was used to detect GFLV in 90 field samples,and the results of the RT-LAMP were in accord with RT-PCR method.The establishment of this method has the characteristics of quick result,convenience,strong specificity and high sensitivity,which is suitable for the rapid screening of the GFLV in customs quarantine,plant protection stations laboratories and other institutions.

关 键 词：葡萄扇叶病毒 反转录环介导等温扩增技术 反转录-聚合酶链式反应 检测 

分 类 号：S41[农业科学—植物保护] Q945.8[农业科学—植物病理学]