检测结核分枝杆菌CFP10-ESAT-6分泌蛋白双抗体夹心ELISA的建立  

Establishment a dual antibody sandwich ELISA to detect secretion protein of Mycobacterium tuberculosis

作　　者：唐宇龙[1,2] 刘湘新[3] 杨华[1] 柳彬[3] 胡忠义[1] 

机构地区：[1]上海市结核重点实验室 [2]湖南农业大学动物医学院,湖南长沙410128 [3]湖南农业大学动物医学院

出　　处：《中国兽医学报》2008年第9期1040-1042,1056,共4页Chinese Journal of Veterinary Science

基　　金：上海市科委重点专项基金资助项目(05DZ22320)

摘　　要：用重组蛋白CFP10-ESAT-6免疫新西兰大白兔,通过ELISA方法检测抗体效价水平;高效价抗体用DEAE-32阴离子交换柱纯化后,一部分抗体为包被所用,另一部分抗体用辣根过氧化物酶标记后,作为检测用,建立双抗体夹心法,进行了初步鉴定,确定该方法的可行性。结果表明:抗CFP10-ESAT-6多克隆抗体的效价在1∶16000稀释后D450值基本都达到1.0左右,在纯化、酶标记后,分别以1∶5000为包被工作浓度,1∶3000为酶标工作浓度时,能较高特异性检测结核杆菌培养上清分泌的CFP10-ESAT-6抗原。A recombinanti CFP10-ESAT-6 protein was prepared to immunize New Zealand rabbits.Titer of antibody was tested through ELISA,high titer of antibody was purified by DEAE-32 anion exchange pillar.Some antibodies were used for coating and another part of the antibodies enzyme-labelled for testing to evaluate the feasibility of the method.Results showed when CFP10-ESAT-6 multi-cloning antibody was diluted to 1∶16 000,the high titer of antibody basically reached 1.0% at D450,and after purification and enzyme-labe...

关 键 词：结核分枝杆菌 CFP10-ESAT-6 ELISA 

分 类 号：S854.43[农业科学—临床兽医学]