氯霉素间接竞争ELISA(ciELISA)检测方法的建立

Establishment of the Standard Curve of Detecting the Residues of CAP in Food of Animal Origin by Indirect Competitive Enzyme-linked Immunosorbent Assay

以人工合成的氯霉素－牛血清白蛋白（ＣＡＰ－ＢＳＡ）为包被抗原，氯霉素（Ｃｈｌｏｒａｍｐｈｅｎｉｃｏｌ，ＣＡＰ）为竞争的半抗原，两者与一定量的抗ＣＡＰ单抗（ＣＡＰ－ＭｃＡｂ）反应。实验结果表明，理想的包被抗原浓度为１．２５μｇ／ｍｌ，抗ＣＡＰ－ＭｃＡｂ工作浓度为１：１２０００，酶标二抗工作浓度为 １： ５０００，可测最适范围为 １ｎｇ／ｍｌ－１００ｎｇ／ｍｌ，最小检测量为０．１ｎｇ／ｍｌ，批内和批间变异系数分别为３． ６２％和 ５． １９％。得到回归方程 ｙ ＝１．２７３０－ ０．６７４５ｘ（ｒ２＝ ０． ９７７９）和标准曲线，从而建立了快速定量测定 ＣＡＰ含量的间接竞争酶联免疫吸附试验（ＥＬＩＳＡ）。整个测定时间为６小时。

CAP-BSA was man-made coating antigen. The CAP hapten was an competitor to CAP-BSA. They could react to the limited amount of monoclonal antibody against CAP.The result of experiment that the optimal concentration of the coating antigen, monoclonal antibody against CAP and sheep anti-mice IgG were 1 .25ug/ml, 1: 12000 and 1: 5000 respectively. Quantization of the CAP was linear from Ing/ml -100ug/ml.with the detection limit being 0. 1ng/ml.The coefficients of variation of intra-assay and inter-assay we...