黄瓜花叶病毒NASBA检测技术的建立  

Establishment of nucleic acid sequence based amplification system for detection of Cucumber mosaic virus

作　　者：王小明[1,2] 王健华[1] 冯团诚[1] 张雨良[1] 吴育鹏[1,2] 刘志昕[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所农业部热带作物生物技术重点开放实验室,海口571101 [2]海南大学环境与植物保护学院,儋州571737

出　　处：《植物病理学报》2011年第2期139-145,共7页Acta Phytopathologica Sinica

基　　金：国家科技支撑计划课题(2007BAD48B01);中央级公益性科研院所基本科研业务费专项(ITBBZD0754)

摘　　要：以香蕉花叶病病样为材料,初步建立了黄瓜花叶病毒核酸序列依赖性扩增(Nucleic acid sequence based amplifica-tion,NASBA)的检测技术。通过以香蕉叶片总RNA为模板,在黄瓜花叶病毒(Cucumber mosaic virus,CMV)亚组ⅠRNA 2高保守区设计特异引物,进行NASBA反应,经5%琼脂糖凝胶电泳检测,阳性样品中出现了预期大小为310 bp的条带,而阴性和空白对照中均未出现。并对11份香蕉样品分别进行NASBA反应,并经过斑点杂交验证与RT-PCR检测比较,两者的检测结果一致,灵敏度相当,检出限量可达100 pg。The diseased samples of banana infected with CMV（Cucumber mosaic virus） were collected and a nucleic acid sequence based amplification（NASBA） detection system was developed.The specific primers were designed according to the high conserved region of RNA 2 sequence of CMV subgroupⅠ.The 310 bp specific amplification product was obtained in positive sample determined with 5% agarose gel electrophoresis.The 11 different banana samples were analyzed with NASBA and RT-PCR to find out reliability of the me-thod,the former was demonstrated by dot blot with digoxigenin-labeled probe and had the similar results with RT-PCR.Further,the sensitivities were also identical and was equal to 100 pg.

关 键 词：香蕉 黄瓜花叶病毒 核酸序列依赖性扩增 斑点杂交 

分 类 号：S436.421[农业科学—农业昆虫与害虫防治]