基于RPA-EXO的马铃薯金线虫快速检测技术研究  

Development of RPA-EXO assay for detection of potato golden nematode Globodera rostochiensis

作　　者：焦莉苹 冯自洋 雷荣[1] 葛建军[1] 陈井生 王旭[1] 刘大伟[2] Jiao Liping;Feng Ziyang;Lei Rong;Ge Jianjun;Chen Jingsheng;Wang Xu;Liu Dawei(Chinese Academy of Inspection and Quarantine,Beijing 100176,China;Northeast Agricultural University;Chongqing Three Gorges University)

机构地区：[1]中国检验检疫科学研究院,北京100716 [2]东北农业大学 [3]重庆三峡学院

出　　处：《植物检疫》2024年第5期42-46,共5页Plant Quarantine

基　　金：中国检科院基本科研项目(2022JK40);海南省科技专项资助重点研发项目(ZDYF2023XDNY079)。

摘　　要：马铃薯孢囊线虫是国际上重要的检疫性有害生物,可危害马铃薯并造成重大经济损失。本研究基于马铃薯金线虫(Globodera rostochiensis)的ITS序列设计RPA特异性引物和EXO荧光探针,建立了马铃薯金线虫的RPA-EXO检测方法。经过RPA条件优化,RPA特异性引物检测的最佳条件为38℃,30 min。本研究建立的RPA-EXO检测方法可从5个其他植物线虫种群中特异检测出马铃薯金线虫,灵敏度可达到10-3单孢囊。马铃薯金线虫RPA-EXO快速检测方法操作简便、反应灵敏,为基层工作人员提供了一种快速检测马铃薯金线虫的方法。Potato cyst nematode(PCN)is an important quarantine pest in the world and causes major loss in potato crops.In this study,RPA(Recombinase polymerase amplification)specific primers and fluorescent probe were designed based on the ITS sequence of Globodera rostochiensis,and an RPA-EXO detection method for G.rostochiensis was established.By optimizing the reaction time and temperature of RPA,the optimal conditions for RPA detection method were 38℃for 30 min.This assay can distinguish G.rostochiensis from five other plant nematode populations.The sensitivity of RPA-EXO detection method for G.rostochiensis can be achieved 1o-3 single cyst DNA.This is the method for diagnosis G.rostochiensis using RPA-EXO,it's a rapid,convenient and sensitive detection assay.It provides a new rapid method for the detection of G.rostochiensis for grassroots personnel.

关 键 词：马铃薯金线虫 RPA-EXO 检测 

分 类 号：S41[农业科学—植物保护]