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-微生物学报.PDFVIP

微生物学通报 OCT 20, 2009, 36(10): 1614~1617 Microbiology © 2009 by Institute of Microbiology, CAS tongbao@ 生物实验室 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 1 1 1 1 2 2 1* 董晓辉 孟春梅 黎军英 吴建祥 荣 松 张 超 洪 健 (1. 310029) (2 311231) 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(Tissue blot-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; Tissue blot-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-ELISA 1:320 稀释结果相当, 前 8 次印迹均可以得到满 意的检测效果。Tissue blot-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。 关键词: 建兰花叶病毒, 免疫斑点法, 组织印迹法, 单克隆抗体 Detection of CymMV in Orchids Tissue by Dot-ELISA and Tissue Blot-ELISA with Monoclonal Antibody DONG Xiao-Hui1 MENG Chun-Mei1 LI Jun-Ying1 WU Jian-Xiang1 RONG Song2 ZHANG Chao2 HONG Jian1* (1. Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang 310029, China) (2. Zhejiang Chuanhua Biological Technology Co. Ltd., Hangzhou, Zhejiang 311231, China) Abstract: 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. Compariso