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[目的]建立检测食品中脱氧雪腐镰刀菌烯醇(DON)快速灵敏化学发光酶免疫方法。[方法]采用棋盘滴定法确定化学发光检测DON最佳工作条件(抗原包被浓度、包被量、一抗和二抗的工作浓度、线性范围和检出限等),建立化学发光检测DON的方法并对实际样品进行检测,同时将检测结果与LC-MS/MS结果进行对比。[结果]经多次实验,化学发光检测DON的工作条件为:DON-BSA最佳包被浓度为0.1μg/ml,包被量为100μl/孔,抗DON单克隆抗体的最佳工作浓度为1:10000,标准曲线线性范围5～200ng/ml,50%抑制浓度为1.02ng/ml,DON的最低检出浓度为0.05ng/ml,样品中DON的最低检出量为1μg/kg,在100～2000μg/kg添加水平的回收率范围为81%～125%。用所建方法对23个食品样品进行了分析,检测结果与LC-MS/MS结果高度相关(r2=0.98)。[结论]所建方法准确、灵敏,适用于食品中DON污染水平的批量筛选。
[Objective] To establish a rapid and sensitive chemiluminescent enzyme immunoassay for detecting deoxynivalenol (DON) in food. [Method] The chemostat titration method was used to determine the optimum conditions of DON detection by chemiluminescence (antigen concentration, coating volume, working concentration, linear range and detection limit of primary and secondary antibodies, etc.) Methods The actual samples were tested, and the results were compared with the results of LC-MS / MS. [Result] After several experiments, the working conditions of detecting DON by chemiluminescence were as follows: the best coating concentration of DON-BSA was 0.1μg / ml, the coating amount was 100μl / well, the best working concentration of anti-DON monoclonal antibody 1: 10000, the standard curve linear range of 5 ~ 200ng / ml, 50% inhibitory concentration of 1.02ng / ml, DON minimum detectable concentration of 0.05ng / ml, the minimum detectable amount of DON in the sample 1μg / kg, The recoveries ranged from 81% to 125% at 100-2000 μg / kg. Twenty-three food samples were analyzed using the established method, and the results were highly correlated with LC-MS / MS (r2 = 0.98). [Conclusion] The proposed method was accurate and sensitive and suitable for batch screening of DON contamination in food.