用滚环扩增技术检测双生病毒  

Rolling circle amplification detection for geminiviruses

作　　者：余玲玲[1] 钱亚娟[1] 周雪平[1] 

机构地区：[1]浙江大学生物技术研究所,杭州310029

出　　处：《植物病理学报》2008年第6期570-575,共6页Acta Phytopathologica Sinica

基　　金：国家自然科学基金资助项目(30770092)

摘　　要：本文报道了利用滚环扩增技术(rolling circle amplification,RCA)放大双生病毒的靶核酸信号,进而利用限制性内切酶片段长度多态性(RFLP)检测不同类型双生病毒的新技术体系。研究结果表明,利用RCA-RFLP对于已知的单组份和双组份以及带有卫星的菜豆金色花叶病毒属病毒都能够有效检测,对采自浙江杭州地区的田间样品进行RCA-RFLP检测的结果与利用简并引物PA和PB进行PCR扩增的结果相吻合。相比传统的PCR检测方法,用RCA-RFLP体系检测双生病毒灵敏性更高,操作更简便。A new method based on rolling circle amplification and restriction fragment length polymorphism （RCA-RFLP） to detect viral DNA components associated with geminiviruses is reported. The results demonstrated the high efficiency of RCA-RFLP in characterizing different geminiviruses, including monopartite or bipartite geminiviruses and their associated DNAβ. Further analysis for field samples from Hangzhou, Zhejiang province, suggested that RCA-RFLP could be successfully used to diagnose geminiviruses in natural infected plants. In addition, RCA-RFLP is more convenient and sensitive for detection geminiviruses than PCR.

关 键 词：滚环扩增 限制性内切酶片段长度多态性 检测 双生病毒 

分 类 号：S432.41[农业科学—植物病理学]