免疫凝聚试纸条和TaqMan探针实时荧光PCR检测西瓜细菌性果斑病菌比较研究  

Comparison of immunoStrip and real-time fluorescent PCR (TaqMan) for detection of Acidovorax avenae subsp. citrulli, the causal agent of bacterial fruit blotch of watermelon

作　　者：冯建军[1] 许勇[2] 李健强[1] Norm W. SCHAAD 

机构地区：[1]中国农业大学植物病理学系,北京100094 [2]国家蔬菜工程技术研究中心,北京100089 [3]美国农业部农业研究中心外来生物与杂草研究所,马里兰州21702

出　　处：《植物病理学报》2006年第2期102-108,共7页Acta Phytopathologica Sinica

基　　金：农业部"九四八"项目(201024);科技部农业成果转化基金资助项目(03EFN211100008);教育部"新世纪优秀人才支持计划"项目(NCET-04-0128)

摘　　要：以西瓜细菌性果斑病菌（Acidovorax avenae subsp．citrulli）菌悬液和田间采集的病组织为试材，研究了免疫凝聚试纸条和实时荧光PCR技术检测的灵敏度和适应性。结果表明，免疫凝聚试纸条检测灵敏度为10^6 cfu／mL，具有简便、快速、易操作特点，适用于田间快速检测和病害诊断；TaqMan探针实时荧光PCR检测灵敏度达10^3～4 cfu／mL，比传统PCR检测灵敏度（10^5 cfu／mL）提高了10～100倍，且不需要琼脂糖凝胶电泳、溴化乙锭染色和Southern杂交。但需要昂贵的仪器和试剂，适用于室内检测及相关研究。To assess the threshold and efficiency of immunoStrip and real-time fluorescent PCR for detection of Acidovorax avenae subsp, citrulli in cell suspension from pure culture and in extracts from naturally infested or infected tissues, a comparative study was conducted. The results showed that immunoStrip testing, with threshold 10^6 cfu/mL, was simple, fast and easy to handle, which adapted for fast detection and diagnosis in field. Compared with classical-PCR （threshold 10^5 cfu/mL）, real-time fluorescent PCR （TaqMan） didn＇t require agarose gel electrophoresis, ethidium bromide staining, and Southern blotting. Moreover, it had a much higher sensitivity, accounting for 10^3～4 cfu/mL, but required expensive instrument, and was adapted to lab testing and basal research.

关 键 词：西瓜 细菌性果斑病菌 免疫凝聚试纸检测 TAQMAN探针 实时荧光PCR检测 

分 类 号：S436.5[农业科学—农业昆虫与害虫防治]