畜牧与兽医2012，Vol.44Issue(6)：10-16,7.
双抗体夹心ELISA检测水牛γ-干扰素方法的建立及初步应用
Establishment of double antibody sandwich ELISA for water buffalo interferon-gamma and its application in tuberculosis diagnosis
摘要
Abstract
Double antibody sandwich ELISA (DAS-EIiSA) for IFN-γ in water buffalo was developed. In this method, two mice ascites McAbs against water buffalo intetferon-γ were purified by ammonium sulfate precipitation (AS), and McAbs 1E4 was used as a capture antibody and HRP-labeled McAbs 4A11 as a detection antibody. The optimal reaction conditions for the assay were determined. The result indicated that the optimal concentration of capture antibody was 625ng/mL, the working concentration of HRP-labeled 4A11 was 1 : 100, and the detection limit of the sandwich ELISA for IFN-γ was 25ng/mL. Compared with tubculin skin test (TST), the diagnosis specificity, sensitivity and coincidence test of DAS-ELISA were 62% , 87% and 75% , respectively. Therefore, this DAS-ELJSA assay will be a useful tool in the bovine tuberculosis diagnosis in water buffalo.关键词
结核病/IFN-γ/ELISA/ESAT-6/CFP-10/单克隆抗体Key words
water buffalo/ IFN-γ/ ELISA/ ESAT-6/CFP-10/ monoclonal antibody (McAb)分类
农业科技引用本文复制引用
徐贤坤,熊毅,刘棋,曾宪垠,龙剑明..双抗体夹心ELISA检测水牛γ-干扰素方法的建立及初步应用[J].畜牧与兽医,2012,44(6):10-16,7.基金项目
广西科技厅科技攻关项目(桂科攻0719004-3F). （桂科攻0719004-3F）
