LAMP方法检测豌豆细菌性疫病菌  

Detection of Pseudomonas syringae pv.pisi by LAMP(Loop-mediated isothermal amplification)

作　　者：封立平[1] 尼秀媚[1] 厉艳[1] 吴兴海[1] 纪瑛[1] 甘琴华[1] 

机构地区：[1]山东出入境检验检疫局,山东青岛266002

出　　处：《植物检疫》2013年第3期80-84,共5页Plant Quarantine

基　　金：质检总局科研项目(2009IK254)

摘　　要：本文应用环介导等温核酸扩增技术(LAMP)对豌豆细菌性疫病菌的肽相关脂蛋白(oprL)特异性基因进行扩增,采用2对引物识别目的片段的6个特异性区域,60 oC恒温1h完成扩增,设计的引物与假单胞属的近缘菌丁香假单胞菌丁香致病变种、绿黄假单胞菌、丁香假单胞菌大豆致病变种、丁香假单胞菌菜豆致病变种都没有扩增反应,表现了较好的特异性。该LAMP检测方法与普通PCR相比灵敏度高100倍。同时,LAMP技术的检测程序便捷,所需设备简单,其结果可以通过肉眼观察来判断,比常规PCR能更早得到反应结果,因此该技术在该病菌的口岸检疫方面拥有更大的优势。Detection of oprL DNA of Pseudomonas syringae pv.pisi by a loop-mediated isothermal amplification （LAMP） was evaluated in laboratory assays. LAMP amplifies target DNA rapidly （1 hour）, isothermally （60~C） and with high-specificity based on two primers designed to recognize six independent sequences of target DNA. The LAMP primers did not react with suspensions of Pseudomonas syringae pv. syringae, Pseudomonas viridijqava, Pseudomonas syringae pv. glycinea, Pseudomonas syringae pv. phaseolicola. The sensitivity of this LAMP assay is higher than PCR. Detecting oprL DNA with LAMP has simpler instrumental requirement and more convenient test procedures, which need shorter time than PCR. In addition,the results can be judged by macroscopic observation. So the technique may have more advantages in port quarantine.

关 键 词：豌豆细菌性疫病菌 LAMP oprL 检测 

分 类 号：S41-31[农业科学—植物保护]