氯霉素间接竞争ELISA检测方法的建立

Development of Ci-ELISA detection method of Chloramphenicol

为了对动物性食品中氯霉素(CAP)进行快速检测,以CAP与卵清蛋白(OVA)偶联合成的CAP-OVA为包被抗原,CAP为竞争的半抗原,通过优化反应条件,建立间接竞争ELISA检测方法.结果显示:最佳包被抗原质量浓度为0.25μg/mL,单克隆抗体稀释倍数为1∶32000,酶标二抗稀释倍数为1∶4000,最低检测限为1.01 ng/mL,批内和批间变异系数分别为3.80%和6.30%,建立的回归方程为y=-10.7x+66.042(R2=0.9969),建立了快速检测氯霉素残留的间接竞争ELISA法.

In order to rapidly detect Chloramphenicol(CAP)in animal derived food,CAP was coupled with ovalbumin(OVA)to synthesize coating antigen(CAP-OVA),and CAP was the competitor to CAP-OVA.The indirect competitive ELISA assay(ci-ELISA)was established by optimizing the reactive conditions.The experimental results showed that the optimal working concentration of the coated antigen was 0.25μg/mL,and the optimal dilution ratio of the monoclonal antibody was 1∶32000.The optimal working concentration of the enzyme-labeled secondary antibody is 1∶4000,and the minimum detection limit is 1.01 ng/mL.The intra-assay and inter-assay coefficients of variation are 3.80% and 6.30%,respectively.The established regression equation is y=-10.7x+66.042(R2=0.9969).Thus,an indirect competitive ELISA method for rapid detection of chloramphenicol residues was established.