菌物学报2025，Vol.44Issue(6)：153-161,9.DOI:10.13346/j.mycosystema.240306
基于RPA-LFD的小麦矮腥黑粉菌可视化快速检测方法的建立
Establishment of rapid detection method of wheat dwarf bunt pathogen Tilletia controversa based on RPA-LFD
摘要
Abstract
Wheat dwarf smut is one of the most harmful systematic infections caused by Tilletia controversa(TCK),which is also an important international quarantine disease of wheat.The accurate and rapid detection of the pathogen is helpful to prevent diseases from being introduced into China along with the port cargo transport.Based on the OR gene specific sequence of TCK,this study designed recombinant polymerase amplification(RPA)and side-flow chromatography strip(LFD)primers TCKRPA3F/3R and probes nfo prode,and the rapid visualization RPA-LFD detection system for TCK was developed.The RPA-LFD system could detect TCK within 10 minutes at 38℃.The minimum detection limits for TCK genomic DNA concentration and recombinant plasmid copy number constructed with targeted specific fragments were 1 pg/μL and 102 copies/μL,respectively,and the minimum limit for the number of teliospores of TCK was 50.Compared with common PCR method,the sensitivity of this method is much higher.Moreover,the detection results can be visually read through the test strip,making the detection easy to operate.This method is suitable for the inspection and quarantine requirements of TCK at basic customs.关键词
小麦矮腥黑粉菌/重组酶聚合酶扩增技术/侧流层析试纸条/可视化快速检测Key words
Tilletia controversa/recombinase polymerase amplification/lateral flow dipstick/visual rapid detection引用本文复制引用
黄洁,崇钰,李妍,郭文静,王保通,李强..基于RPA-LFD的小麦矮腥黑粉菌可视化快速检测方法的建立[J].菌物学报,2025,44(6):153-161,9.基金项目
科技部对发展中国家援助项目(KY202002018)This work was supported by the Ministry of Science and Technology's Aid Projects for Developing Countries(KY202002018). （KY202002018）
