夹心ELISA检测牛γ-干扰素方法的建立及应用

摘要

To develop a sandwich ELISA for detection of bovine interferon γ (BoIFN-γ), 3 monoclonal antibodies against BoIFN-γ were tested as capture antibodies. All of 3 monoclonal antibodies recognized BoIFN-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 BoIFN-γ in bovine tuberculosis disease.%为建立检测牛γ-干扰素（BoIFN-γ）的双夹心ELISA方法，将前期获得的针对BoIFN-γ的单克隆抗体作为捕捉抗体进行夹心ELISA检测天然BoIFN-γ，结果表明单克隆抗体BoIFN-γ-6E5、BoIFN-γ-3E1、BoIFN-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BoIFN-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。