| 氯霉素间接竞争ELISA(ciELISA)检测方法的建立 |
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| 引用本文： | 周斌,石德时,覃雅丽,王桂枝.氯霉素间接竞争ELISA(ciELISA)检测方法的建立[J].中国预防兽医学报,2000(Z1). |
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| 作者姓名： | 周斌 石德时 覃雅丽 王桂枝 |
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| 作者单位： | 华中农业大学畜牧兽医学院!武汉 430070 |
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| 基金项目： | 湖北省自然科学基金资助项目!（项目编号：96J055） |
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| 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（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。
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| 关 键 词： | 氯霉素残留 ciELISA 标准曲线 |
Establishment of the Standard Curve of Detecting the Residues of CAP in Food of Animal Origin by Indirect Competitive Enzyme-linked Immunosorbent Assay |
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| ZHOU Bin,SHI Deshi,QIN Yali,WANG Guizhi.Establishment of the Standard Curve of Detecting the Residues of CAP in Food of Animal Origin by Indirect Competitive Enzyme-linked Immunosorbent Assay[J].Chinese Journal of Preventive Veterinary Medicine,2000(Z1). |
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| Authors: | ZHOU Bin SHI Deshi QIN Yali WANG Guizhi |
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| Abstract: | 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 were 3 .62% and 5. 19% respectively.Then,the regression equation was y = 1 .2730 - 0.6745x(r2= 0 .9779) and there was a standard curve. SO indirect competitive enzyme-linked immunosorbent assay was established to detect rapidly the residues of CAP in food of animal origin. The whole assay could be completed within 6 hours. |
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| Keywords: | Residues of CAP ciELISA The standard curve |
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