恩诺沙星化学发光免疫分析试剂盒的研制
Development of an indirect competitive chemiluminescence enzyme immunoassay kit for detecting of enrofloxacin residue
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摘要: 主要研制一种基于鲁米诺-辣根过氧化物酶体系的化学发光免疫分析试剂盒,用于检测食品中的痕量恩诺沙星残留。反应条件优化结果为:最佳包被抗原浓度为0.5ng/mL,最佳抗体稀释倍数为1:2000,药物稀释液为0.01mol/L的PBST溶液,一抗反应时间为30min。研究结果显示:本文研制的试剂盒的回归曲线方程是y=-22842x+172936,检测线性范围是0.024～2.76ng/mL,IC50为0.106ng/mL,对蜂蜜、牛奶、鱼肉和虾肉样本的添加回收率在96%～116%、90%～97%、92%～112%、86%～112%之间,批内和批间变异系数均小于15%,试剂盒在4℃下有效期为180d。Abstract: A HRP-Luminol-H2O2 system based indirect competitive chemiluminescence enzyme immunoassay (CLEIA) kit for determination of enrofloxacin (ENR) residues was developed.The CLEIA conditions were optimized.The optimal concentration of coating antigen was 0.5ng/mL and the antibody dilution was 1:2000.Using PBST as assay buffer, the incubation time for the primary antibody was 30min.Under the optimal conditions, the standard curve was developed.The regression equation for ENR was y=-22842x+172936 with a linear detection ranging from 0.024 to 2.76ng/mL.The IC50 was 0.106ng/mL.The averaged recoveries of ENR from spiked honey, milk, shrimp and fish samples were 96%～116%, 90%～97%, 92%～112%and 86%～112%, respectively.The coefficient of variation of intra-assay and inter-assay was less than 15%.The developed kit can be stored at 4℃ for 180 days.
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