单抗免疫斑点法和组织印迹法检测百合无症病毒.PDFVIP

微生物学通报. 2006, 33(2): 25-29 单抗免疫斑点法和组织印迹法检测百合无症病毒* 1 1 1 * * 1 2 李燕宏 吴建祥 洪 健 周雪平 叶美琴 1 2 （浙江大学生物技术研究所 杭州 310029 ） （浙江丽水市农业局 丽水 323000 ） 摘要：应用抗百合无症病毒（Lily symptomless virus ，LSV ）的单克隆抗体，建立了快速检测田间样品的免疫斑 点法（Dot-ELISA ）和组织印迹法（Tissue blot-ELISA ）体系。LSV 单抗稀释2000 倍时，Dot-ELISA 中病叶粗 汁液可被检出的最大稀释度为1:640 。Tissue blot-ELISA 中样品一次平切后第1 次印迹与Dot-ELISA 样品 1:40 稀释的结果相当，前4 次印迹均可获得满意的显色效果。常规Tissue blot-ELISA 的灵敏度低于Dot-ELISA，但 用丙酮处理点过样的硝酸纤维素膜后，二者的灵敏度相当，且Tissue blot-ELISA 操作更简便，适合田间大量样 品的快速检测。 关键词：百合无症病毒；单克隆抗体；免疫斑点法；组织印迹法 中图分类号：Q93 文献标识码：A DETECTION OF LSV IN LILY TISSUE BY DOT-ELISA AND TISSUE BLOT-ELISA WITH MONOCLONAL ANTIBODY* 1 1 1 * * 1 2 LI Yan-Hong WU Jian-Xiang HONG Jian ZHOU Xue-Ping YE Mei-Qing (Institute of Biotechnology , Zhejiang University , Hangzhou 310029 ) 1 (Agriculture Bureau of Lishui , Lishui Zhejiang 323000) 2 Abstract: Dot-ELISA and tissue blot-ELISA with monoclonal antibody to detect Lily symptomless virus (LSV) were established. 2000-dilution LSV monoclonal antibody could be adopted to detect LSV infecting leaf extract within a dilution limitation of 1:640. LSV could still be detected in the forth print by tissue blot-ELISA . Similar results were shown between the first print by tissue blot-ELISA and 40-dilution of infected leaf extract by dot-ELISA. Comparison tests showed that the commonly used tissue blot-ELISA was less sensitive than dot-ELISA, b