中国兽医科学2012，Vol.42Issue(7)：696-700,5.
鸭疫里氏杆菌双抗体夹心ELISA检测方法的建立
Establishment of a double antibody sandwich ELISA for the detection of Riemerella anatipestifer
摘要
Abstract
The aim of the present study was to develop a rapid and accurate method for the detection of Riemerella anatipestifer based on a double antibody sandwich ELISA. Purified polyclonal antibody against serotype 1 R. anatipestifer was used as a capture antibody, while purified monoclonal antibody 5G7 was used as a detection antibody. The optimal conditions for the ELISA were as follows：polyclonal antibody with 184 ng per well was added to coat overnight,the plate was blocked by 100 mL/L neonatal calf serum for 2 hours at 37℃ ,the incubation times for detection antigen,monoclonal antibody and second labeled an- tibody were 1 hour at 37℃for each,the value of D450nm was obtained after 20 min coloration. This method has no cross-reaction with poultry-origin Pasteurella multocida, Escherichia coli and Salmonella, and its minimum detectable limit was 104 CFU/mL. The sensitivity of the method was 100% ,and the coincidence between bacterial isolation method and the double antibody sandwich ELISA was 85.2M. Therefore,this study provides a rapid, specific and sensitive detection method for R. anatipestifer.关键词
鸭疫里氏杆菌/单克隆抗体/双抗体夹心ELISAKey words
Riemerella anatipestifer/monoclonal antibody/double antibody sandwich ELISA分类
农业科技引用本文复制引用
陈素娟,陈冰,李鑫,高以明,彭大新,刘秀梵..鸭疫里氏杆菌双抗体夹心ELISA检测方法的建立[J].中国兽医科学,2012,42(7):696-700,5.基金项目
公益性行业（农业）科研专项 （）
江苏省检验检疫局科研项目计划项目 （）
江苏省青蓝工程项目 （）
教育部“长江学者和创新团队发展计划”创新团队项目 （）
