玉米细菌性枯萎病菌LAMP检测方法的建立  

LAMP for the detection of Pantoea stewartii subsp. stewartii

作　　者：袁钧[1] 高文娜[2] 汪万春[2] 廖晓兰[1] 

机构地区：[1]湖南农业大学生物安全科技学院,湖南长沙410128 [2]北京出入境检验检疫局技术中心

出　　处：《植物检疫》2013年第5期61-65,共5页Plant Quarantine

基　　金：国家"十二五"重大科技支撑计划(2012BAK11B02-1)

摘　　要：利用玉米细菌性枯萎病菌基因组DNA特有的保守区域设计LAMP(loop-mediated isothermal amplification)引物,通过优化其反应条件,建立了玉米细菌性枯萎病菌的LAMP检测体系。以多种参比菌DNA为模板对LAMP检测体系的特异性进行了验证,利用玉米细菌性枯萎病菌DNA溶液和菌液梯度稀释液对LAMP检测体系的灵敏度进行了验证。在特异性测试中,LAMP检测体系仅对玉米细菌性枯萎病菌进行扩增,对非靶标菌不产生扩增。LAMP检测体系DNA和菌体检测灵敏度分别达到了0.858×10-4ng/uL和45 CFU/mL。为玉米细菌性枯萎病菌的检测提供了一种新的、简便的、快速的检测手段。The specific region of the genome of Pantoea stewartii subsp.stewartii was used for the design of LAMP primers,the LAMP detection system were established by optimization of reaction conditions.Specificity of the system was verified using a series of reference strains,sensitivity of the system was verified using a series of gradient dilution of DNA solution of the Pantoea stewartii subsp.stewartii and the bacterial suspension.In the specificity test,LAMP system showed high and identical specificity.DNA and bacterial suspension sensitivity of LAMP system reached 0.858×10-4ng/uL and 45 CFU/mL respectively.LAMP system provide a new,fast and easy way for the detection of Pantoea stewartii subsp.stewartii.

关 键 词：玉米细菌性枯萎病菌 环介导等温扩增 检测 

分 类 号：S435.13[农业科学—农业昆虫与害虫防治]