氯霉素酶联免疫检测方法的研究

An Enzyme Immunoassay for the Determination of Chloramphenicol

通过活化琥珀氯霉素的羧基使其与蛋白联结合成了免疫抗原 ,并制备了氯霉素的多克隆抗体 ,建立了氯霉素直接竞争酶联免疫检测方法 ,检测时间在 1h之内。抗体的ELISA效价达 3.2× 10 5以上 ,I50 为 12 .9ng ml,检测范围为 0 .2 4ng ml- 6 78.8ng ml。抗体与琥珀氯霉素的交叉反应率为 174 .5 4 % ,与其它抗生素的交叉反应率均小于 0 .0 1%。牛奶样品中平均添加回收率为10 6 .5 6 %。所建立的氯霉素酶联免疫检测方法达到了我国牛奶氯霉素残留限量 0 .2 5ng ml的要求 。

A direct competitive enzyme immunoassay for the quantitative analysis of CAP is described.Polyclonal antibodies(PAbs) with high affinity to chloramphenicol were prepared from rabbits immunized with CAP succinate conjugated to protein.The titer of antiserum was 3.2×105,I_(50) was 12.9 ng/ml.CAP could be quantitated in the range 0.24-678.8 ng/ml.The cross-reactivity of PAbs with seven antibiotics were <0.01%.Average recovery of CAP from milk was 106.56%.The detection limit was 0.24ng/ml.belowed maximum residue limits(MRLs) established by the Peoples Republic of China (PRC).This research will be useful for the development of anti-CAP EIA Kit.