单抗免疫斑点法和组织印迹法检测侵染蝴蝶兰的建兰花叶病毒  

Detection of CymMV in Orchids Tissue by Dot-ELISA and Tissue Blot-ELISA with Monoclonal Antibody

作　　者：董晓辉[1] 孟春梅[1] 黎军英[1] 吴建祥[1] 荣松 张超 洪健[1] 

机构地区：[1]浙江大学生物技术研究所,浙江杭州310029 [2]浙江传化生物技术有限公司,浙江杭州311231

出　　处：《微生物学通报》2009年第10期1614-1617,共4页Microbiology China

基　　金：杭州市重大科技创新专项(No.20080212A10);浙江省科技厅分析测试科技计划项目(No.2007F70014)

摘　　要：应用抗建兰花叶病毒（CymMV）的单克隆抗体，建立了快速检测蝴蝶兰病样的免疫斑点法（Dot—ELISA）和组织印迹法（Tissueblot—ELISA）。CymMV单抗稀释8000倍时．Dot—ELISA可检出病毒粗汁液的最大稀释度为1：10240；Tissueblot—ELISA中样品1次平切后1～5次印迹与Dot—ELISA样品1：80稀释结果相当，6-8次印迹与Dot—ELISA1：320稀释结果相当，前8次印迹均可以得到满意的检测效果。Tissueblot—ELISA的灵敏度略低于Dot—ELISA，使用单抗的结果比多抗特异性强，操作更为简便，适合大量兰花样品的快速检测。Dot-ELISA and tissue blot-ELISA with monoclonal antibody to detect Cymbidium mosaic virus （CymMV） were established. 8000-dilution CymMV monoclonal antibody could be adopted to detect CymMV infecting leaf extract within a dilution limitation of 1：10240. CymMV could still be detected in the eighth print by tissue blot-ELISA. Similar results were shown between the fifth print by tissue blot-ELISA and 80-dilution of infected leaf extract by dot-ELISA. Comparison tests showed that the commonly used tissue blot-ELISA was less sensitive than dot-ELISA. It could illustrate that CymMV monoclonal antibody was more specific and sensitive than the polyclonal antibodies. The improved tissue blot-ELISA is more simple than dot-ELISA, and is suitable for fast detection with a large number of samples in the field.

关 键 词：建兰花叶病毒 免疫斑点法 组织印迹法 单克隆抗体 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治] S432.41[农业科学—植物保护]