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.
出处
《植物病理学报》
CAS
CSCD
北大核心
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)
关键词
香蕉
黄瓜花叶病毒
核酸序列依赖性扩增
斑点杂交
banana
Cucumber mosaic virus
nucleic acid sequence based amplification
dot blot
