夹心ELISA检测牛γ-干扰素方法的建立及应用  

DEVELOPMENT AND APPLICATION OF SANDWICH ELISA FOR DETECTING BOVINE INTERFERON GAMMA

作　　者：钱琨[1,2,3,4] 赵巍[2,3] 黄大卢[2,3] 邵红霞[2,3,4] 秦爱建[1,2,3,4] 

机构地区：[1]扬州大学兽医学院,扬州225009 [2]江苏省动物预防医学重点实验室,扬州225009 [3]教育部禽类预防医学重点实验室,扬州225009 [4]江苏省动物重要疫病与人兽共患病防控协同创新中心,扬州225009

出　　处：《中国动物传染病学报》2016年第5期57-62,共6页Chinese Journal of Animal Infectious Diseases

基　　金：江苏省科技支撑计划(BE2013391);江苏省优势学科项目

摘　　要：为建立检测牛γ-干扰素(Bo IFN-γ)的双夹心ELISA方法,将前期获得的针对Bo IFN-γ的单克隆抗体作为捕捉抗体进行夹心ELISA检测天然Bo IFN-γ,结果表明单克隆抗体Bo IFN-γ-6E5、Bo IFN-γ-3E1、Bo IFN-γ-3C2三株能捕捉到天然的牛γ干扰素。经过条件摸索和优化,最终建立了以3E1/HRP-6E5为最佳组合的检测Bo IFN-γ的夹心ELISA方法。用建立的双夹心ELISA检测方法和进口试剂盒同时对66份血样进行检测,结果表明双夹心ELISA方法和进口试剂盒检测结果的符合率达95.45%,说明该方法能有效地对牛结核病γ-干扰素进行检测。To develop a sandwich ELISA for detection of bovine interferon γ（Bo IFN-γ）, 3 monoclonal antibodies against Bo IFN-γ were tested as capture antibodies. All of 3 monoclonal antibodies recognized Bo IFN-γ. The sandwich ELISA method was optimized and monoclonal antibody 3E1 was determined as the capture antibody. Total 66 serum samples from different regions were tested using the sandwich ELISA developed here and a commercial ELISA kit. The agreement between these two methods was 95.45%, suggesting the promising possibility of the sandwich ELISA for effective detection of Bo IFN-γ in bovine tuberculosis disease.

关 键 词：牛γ-干扰素 双夹心ELISA 应用 

分 类 号：S852.4[农业科学—基础兽医学]