快速鉴定草莓枯萎病的LAMP检测技术

Rapid Identification of Fusarium Wilt in Strawberries Using LAMP Detection Technique

【目的】建立草莓枯萎病的镰刀菌快速检测方法。【方法】利用环介导等温扩增技术(LAMP)针对镰刀菌保守区域ITS设计一套LAMP扩增引物,利用Bst DNA聚合酶完成LAMP扩增反应,对7种常见病原菌进行检测,分析LAMP引物的特异性;比较LAMP与传统PCR方法的敏感性。【结果】建立的LAMP法能够特异性的检测尖孢镰刀菌和腐皮镰刀菌,并且检测灵敏度为1.0×10^(-4) ng/μL,是普通PCR检测灵敏度的1000倍。【意义】试验所建立的草莓枯萎病快速LAMP检测方法,具有灵敏度高、快速及简便的优点,可以向基层单位推广使用。

[Objectives]A rapid detection method for Fusarium wilt in strawberries was established.[Methods]A set of LAMP amplification primers were designed for the conserved region of ITS in Fusarium spp.to detect seven common pathogens by using loop-mediated isothermal amplification(LAMP),and LAMP amplification reaction was completed by using Bst DNA polymerase.Also,the specificity of LAMP primer was analyzed,and the sensitivity between LAMP and PCR was compared.[Results]The results showed that LAMP method could detect Fusarium oxysporum and Fusarium solani,with the detection sensitivity of 1.0×10^(-4) ng/μL,which was 1000 times that of conventional PCR.[Significance]The rapid detection method LAMP for Fusarium wilt in strawberries developed in this experiment had the advantages of high sensitivity,rapidity and simplicity,and could be popularized to grass-root units.