番茄溃疡病菌LAMP可视化检测方法的建立

Establishment of a LAMP Method for Detection of Clavibacter michiganensis subsp.michiganensis

以番茄细菌性溃疡病菌等7株病原细菌为检测材料,以特异性基因tom A为检测靶标,设计LAMP引物tom A-107、tom A-8和相应的环引物tom A-107LF21、tom A-107LB21。针对靶标基因进行引物筛选及引物的特异性和灵敏度测试,结果表明:tom A-107能特异性检测出番茄细菌性溃疡病菌,检测灵敏度达5×10^(2)copy/μL,环引物的加成将反应进程缩短三分之一,使整个检测过程控制在50 min内完成,该方法非常适用于Cmm菌株的现场快速检测。

In this study,eight bacterial strains,including Clavibacter michiganensis subsp.michiganensis(Cmm),were used as test materials,with the specific gene tomA as the detection target.LAMP primers tomA-107 and tomA-8,as well as the corresponding loop primers tomA-107LF21 and tomA-107LB21,were designed.The primer specificity and sensitivity tests were conducted for the target gene,and the results showed that primer tomA-107 could specifically detect Cmm.The detection sensitivity of primer tomA-107 reached 5×10^(2)copies/μL for Cmm.The addition of loop primers shortened the reaction process by one third,enabling the entire detection process to be completed within 50 minutes.This method is highly suitable for rapid on-site detection of Cmm bacterial strains.