食品中黄曲霉毒素M_(1)的间接竞争酶联免疫法的建立

Determination Method of Aflatoxin M_(1)in Food by Indirect Competitive Enzyme⁃Linked Immunosorbent Assay

该文建立一种间接竞争酶联免疫法(indirect competitive enzyme-linked immunosorbent assay,ic-ELISA)快速测定食品中黄曲霉毒素M_(1)(aflatoxin M_(1),AFM_(1))的分析方法。AFM_(1)标准品用甲醇稀释,AFM_(1)标准品与AFM_(1)全抗原竞争结合AFM_(1)单克隆抗体,利用3,3′,5,5′-四甲基联苯胺(3,3′,5,5′-tetramethylbenzidine,TMB)单组分显色液显色后,酶标仪测定吸光度。在优化好的条件下,浓度范围为9.743~2.605×10^(3)pg/mL时具有良好的线性关系,相关系数(determination coefficient,R^(2))为0.9927,检出限IC_(10)为3.84 pg/mL,加标回收率为88.43%~105.75%。该方法适用性好、操作简单、灵敏度高,满足食品中AFM_(1)分析检测的需求。

This study established an analytical method for the determination of aflatoxin M_(1)(AFM_(1))in food by indirect competitive enzyme⁃linked immunosorbent assay(ic⁃ELISA).AFM_(1)standard,diluted with methanol,competed with AFM_(1)antigen to bind AFM_(1)monoclonal antibody.The absorbance was measured by a micro⁃plate reader after color development with a single⁃component TMB color⁃developing solution.Under optimized conditions,AFM_(1)had a good linear relationship in the concentration range of 9.743-2.605×10^(3)pg/mL,with the determination coefficient(R^(2))of 0.9927,the limit of detection IC_(10)of 3.84 pg/mL,and the recovery rate of 88.43%-105.75%.This determination method had good applicability,simple operation,and high sensitiv⁃ity,and met the requirements of AFM_(1)analysis and detection in food.