醋酸甲孕酮(MPA)间接竞争ELISA检测方法的建立

Establishment of Indirect Competitive Enzyme-linked Immunosorbent Assay for Detecting the Residues of MPA

将半抗原醋酸甲孕酮（MPA）衍生物与牛血清白蛋白（BSA）偶联后，作为包被原与游离的醋酸甲孕酮竞争抗MPA多克隆抗体，通过该模式建立间接竞争ELISA方法检测MPA的残留量。试验结果表明：理想的包被抗原为0．625μg／ml，抗体的工作浓度为1：8000，标准曲线在1～50ng／ml之间线性关系较好，最低检测限0．1ng／ml，得回归方程y=-1．1479x＋0．9426（r^2=0．9679），批内和批间变异系数分别为5．35％、10．89％。

Coupled with the BSA, hapten MPA, used as a coating antigen, combined with anti- MPA polyclonal antibody with the competition of free MPA. Indirect competition ELISA was set up according to above-mentioned mode to detect the residue level of MPA in the example. The results of the experiment demonstrates that the optimal concentration of the coating antigen was 0.75 btg/ml. Polyclonal antibody against MPA was 1 ： 8000 and the standard curve had a good linearity in the rang of 1-50 ng/ml with detection limit being 0.1 ng/ml. The regression equation y= 1. 1479x＋0. 9426（r^2 =0. 9679） were obtained while the coefficient of variation of intra-assay and extra-assay were 5.35 %, 10.89 % respectively.