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
焦莉苹 1冯自洋 2雷荣 3葛建军 3陈井生 4王旭 3刘大伟5
作者信息
- 1. 中国检验检疫科学研究院 北京 100176;东北农业大学
- 2. 中国检验检疫科学研究院 北京 100176;重庆三峡学院
- 3. 中国检验检疫科学研究院 北京 100176
- 4. 重庆三峡学院
- 5. 东北农业大学
- 折叠
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℃,30min.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.
Abstract
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 flu-orescent 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 10-3 single cyst DNA.This is the method for diagnosis G.ros-tochiensis 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Key words
Globodera rostochiensis/RPA-EXO/detection引用本文复制引用
基金项目
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)
出版年
2024
NETL
NSTL
万方数据