氯霉素竞争ELISA检测方法的研究

Development of MAb-based competitive ELISA for detecting chloramphenicol residues

用抗氯霉素的MAb2G2建立了检测氯霉素的竞争ELISA,对主要影响因素进行了研究,其回归方程为y=-0.3522x+0.5939,相关系数为R2=0.9906,检测线性范围为0.098ng/mL～25ng/mL,半数抑制浓度(IC50)为1.848ng/mL,检测限为0.135ng/mL。与氯霉素琥珀酸钠交叉反应率为143%,与其它结构类似物和常见抗菌素的交叉反应率均小于0.01%。批内和批间变异系数分别为4.98%和9.42%,牛奶样的平均添加回收率为101.31%。所建立的检测氯霉素竞争ELISA法,符合检测氯霉素残留的要求,为氯霉素残留快速检测试剂盒的研制奠定了基础。

A competitive ELISA for detecting chloramphenicol was developed using 2G2 monoclonal antibody against chlor- amphenicol. The ELISA had a regression line ofy=-0.3522x＋0.5939 and correlation coefficient 0.9906. The linear range varied from 0.098 ng/mL to 25 ng/mL, with the IC50 of 1.848 ng/mL and the detection limit of 0.135 ng/mL. It showed 143 % cross-reactivity with CAP succinate sodium and less than 0.01% cross-reactivity with other antimicrobial agents or structurally related compounds. The intra-assay and inter-assay coefficients of variation were 4.980 % and 9.420 %, respectively, with the average recovery of 101.31% in milk. The ELISA method met quality criteria for detecting chloramphenicol residues, which would be useful for development of CAP immunoassay kit in the detection of CAP residues.