氯霉素间接竞争ELISA(ciELISA)检测方法的建立  

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

作　　者：石德时[1] 周斌[1] 覃雅丽[1] 王桂枝[1] 

机构地区：[1]华中农业大学畜牧兽医学院,湖北武汉430070

出　　处：《中国兽医学报》2002年第1期77-79,共3页Chinese Journal of Veterinary Science

基　　金：湖北省自然科学基金资助项目 (96J0 5 5 )

摘　　要：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 (CAP)为竞争半抗原 ,两者与一定量的抗CAP- Mc Ab反应。结果表明 ,理想的包被抗原质量浓度为 1.2 5 mg/ L ,抗 CAP- Mc Ab稀释倍数为 1∶ 12 0 0 0 ,酶标二抗稀释倍数为 1∶ 5 0 0 0 ,最适检测范围为 1～ 10 0 μg/ L,最小检测量为 0 .1μg/ L,批内和批间变异系数分别为 3.6 2 %和 5 .19%。得到回归方程 (y =1.2 730 - 0 .6 74 5 x,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 (ci EL ISA)。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 results of the experiment demonstrates that the optimal concertration of the coating antigen,monoclonal antibody against CAP and sheep anti mice IgG were 1 25 μg/L,1∶ 12 000 and 1∶ 5 000 respectively.Quantitation of the CAP was linear from 1~100 μg/L,with the detection limit being 0 1 μg/L.The coefficients of variation of intra assay and inter assay were 3 62% and 5 19% respectively.A standard curve was obtained,the regression equation was y= 1 273 0 - 0 674 5 x(r 2= 0 977 9 ).SO a indirect competitive enzyme linked immunosorbent assay for CAP was established.The whole assay costs 4 hours.

关 键 词：氯霉素残留 CIELISA 标准曲线 检测方法 动物性食品 

分 类 号：S859.84[农业科学—临床兽医学] TS207.53[农业科学—兽医学]