马铃薯环腐病菌LAMP检测方法的建立  

Development of LAMP method for the detection of Clavibacter michiganensis subsp. sepedonicus.

作　　者：汪万春[1] 袁钧[2] 郑春生[1] 高文娜[1] 

机构地区：[1]北京出入境检验检疫局检验检疫技术中心,北京100026 [2]湖南农业大学植物保护学院

出　　处：《植物检疫》2014年第1期29-32,共4页Plant Quarantine

基　　金：国家"十二五"科技支撑计划(2012BAK11B02)

摘　　要：选择马铃薯环腐病菌(Clavibacter michiganensis subsp.sepedonicus)基因组DNA特有的保守区域(pCSL0067)设计一套LAMP引物,通过优化反应条件,建立了马铃薯环腐病菌LAMP检测体系。利用多种参比菌的DNA为模板对LAMP检测体系特异性进行了验证,利用马铃薯环腐病菌DNA溶液和菌液梯度稀释液对LAMP检测体系的检测灵敏度进行了验证。在特异性试验中,LAMP检测体系仅对马铃薯环腐病菌进行扩增,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0.527×10-3 ng/uL和150 CFU/mL。为马铃薯环腐病菌的检测提供了一种新的、简便、快速的检测手段。The specific region of the genome of Clavibacter michiganensis subsp, sepedonicus （pCSL0067） was used for the design of LAMP primers, and 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 C. michiganensis subsp, sepedonicus and the bacterial suspension. In the specificity test, LAMP system showed high and identical specificity. DNA and bacterial suspension sensitivity of LAMP system reached 0.527 × 10.3 ng/uL and 150 CFU/mL respectively. This study provide a new, fast and easy way for the detection of C.michiganensis subsp, sepedonicus.

关 键 词：马铃薯环腐病菌 环介导等温扩增 检测 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]