| 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 |
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| 引用本文： | 董晓辉,孟春梅,黎军英,吴建祥,荣 松,张 超,洪 健. 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[J]. 微生物学通报, 2009, 36(10): 1614-1617 |
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| 作者姓名： | 董晓辉 孟春梅 黎军英 吴建祥 荣 松 张 超 洪 健 |
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| 作者单位： | 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 |
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| 基金项目： | 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) |
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| 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
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| 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 |
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| DONG Xiao-Hui,MENG Chun-Mei,LI Jun-Ying,WU Jian-Xiang,RONG Song,ZHANG Chao and HONG Jian. Detection of CymMV in Orchids Tissue by Dot-ELISA and Tissue Blot-ELISA with Monoclonal Antibody[J]. Microbiology China, 2009, 36(10): 1614-1617 |
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| Authors: | DONG Xiao-Hui MENG Chun-Mei LI Jun-Ying WU Jian-Xiang RONG Song ZHANG Chao HONG Jian |
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| Affiliation: | Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang 310029, China;Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang 310030, China;Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang 310031, China;Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang 310032, China;Zhejiang Chuanhua Biological Technology Co. Ltd., Hangzhou, Zhejiang 311231, China;Zhejiang Chuanhua Biological Technology Co. Ltd., Hangzhou, Zhejiang 311231, China;Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang 310029, China |
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| 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. 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. |
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| Keywords: | Cymbidium mosaic virus Monoclonal antibody Dot-ELISA Tissue blot-ELISA |
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