一种检测储存喷气燃料中特征真菌的新方法--LAMP-LFD

摘要

LAMP-LFD,loop-mediated isothermal amplification (LAMP)combined with a lateral flow dipstick (LFD), was adopted as a new method for rapid detection of Amorphotheca resinae (A. resinae)in jet fuel,and was establised by designed primers that specifically recognized 18S rRNA gene of Amorphotheca resinae as tearget sequence,with the results of the temperature of 63 ℃ within 35 minutes. Four fungi and A. resinae that usually found in jet fuel were specifically selected as test strains,and only A. resinae showed positive reaction by using the LAMP-LFD method. The sensitivity of A. resinae genomic DNA were 26 fg/ μL. Nine jet fuel samples were tested and six of them were detected with presence of A. resinae by the established LAMP-LFD method. The results demonstrated that LAMP-LFD possessed the potential of rapid detection of A. resinae in jet fuel in view of its short duration,simple handling,obvious specificity and high sensitivity advantages.%利用环介导等温扩增（LAMP）与横向流动试纸快速检测技术（LFD）相结合，以Amorphothe-ca resinae的18S rRNA基因为靶序列，设计1套LAMP 引物，建立了反应温度63℃、时间35 min。用喷气燃料中曾发现的4株真菌和A． resinae作为特异性试验菌株，LAMP-LFD 法检测只有A． resi-nae呈阳性反应。A． resinae 基因组DNA 检测灵敏度为26 fg ／μL；用已建立的LAMP-LFD 方法对9个喷气燃料样品进行检测，其中有6个样品被检出存在A． resinae。结果表明该方法用于快速检测喷气燃料中的A． resinae耗时短、操作简便、特异性强、灵敏度高，有望用于为检测喷气燃料中特征真菌A． resinae的新方法。