直接竞争酶联免疫吸附法检测食品中三聚氰胺残留

Detection of melamine in food by direct competitive enzyme-linked immuno sorbent assay method

中文关键词:  三聚氰胺  酶标抗原  直接竞争酶联免疫吸附法

英文关键词:melamine  enzyme labeled antigen  direct competitive enzyme linked immunosorbent assay

中文摘要:

      目的 采用酶标抗原建立高效、高灵敏的三聚氰胺直接竞争酶联免疫吸附法(direct competitive enzyme linked immunosorbent assay, dc-ELISA)检测食品中三聚氰胺(melamine, MEL)残留。方法 以三聚氰胺单克隆抗体包被作为固相抗体, 辣根过氧化酶(horseradish peroxidase, HRP)标记的抗原与标准品(或样品)中三聚氰胺竞争结合抗体, 建立了直接竞争酶联免疫检测体系, 以三聚氰胺标准品建立标准曲线进行定量。结果 方法的IC50为8.84 μg/L, 灵敏度为0.65 μg/L, 线性范围0.9~35 μg/L; 检测样品的回收率在70%~120%之间, 与三聚氰酸交叉反应率为60%, 其他结构类似物基本无交叉反应。结论 本方法灵敏度高、特异性强, 可以满足实际样品的快速检测需求。

英文摘要:

      Objective To establish a rapid, high sensitive method to detect melamine (MEL) residues in food by direct competitive enzyme linked immunosorbent assay (dc-ELISA). Methods The MEL antigen was labeled by horseradish peroxidase (HRP), monoclonal antibody was bounded on the surface of a micro titer plate, the standards (or the samples) competed with MEL antigen for the antibody binding sites, a dc-ELISA was established, and the standard curve of melamine was prepared for quantitative analysis. Results The IC50 of dc-ELISA was 8.84 μg/L, the limit of detection was 0.65 μg/L, the linear detection ranges were 0.9?35 μg/L, the recoveries of all kinds of samples were range from 70.0% to 120%, and the cross reaction with cyanuric acid was 60%, and there was almost no cross reaction with other drugs with similar construction. Conclusion The established method is sensitive and specific, which can meet the rapid detection demand of actual samples.