酶联免疫吸附法快速检测黄曲霉毒素

Rapid Detection of Aflatoxin by Enzyme-linked Immunosorbent Assay

结合间接竞争反应机制,运用酶联免疫吸附技术(enzyme-linked immunosorbent assay,ELISA),从抗原(antigen,Ag)包被时间、体系反应温度、酶标二抗作用时间、显色时间等主要因素开展快速检测黄曲霉毒素改进方法的研究。研究结果表明适宜的优化条件为:采用Ag包被20 h、反应温度24℃左右、酶标二抗作用时间30 min、底物显色时间15min。通过对饲料等11种样本的检测得出:半抑制浓度IC50<0.1μg/L,添加回收率在67%~116%,灵敏度达到0.03μg/L,线性系数>0.99,板内变异小于5%。检测试验结果良好,表明该改进方法具有可行性。

Combined with indirect competitive response mechanism,enzyme-linked immunosorbent assay(ELISA)was used to study improved methods for rapid detection of aflatoxin from main factors of antigen(Ag)including duration of inclusion,systemic reaction temperature,time of enzyme-linked immunosorbent assay,color development time,etc.The results showed that the optimal conditions were as follows:Ag coating was used for 20 h,the reaction temperature was around 24℃,the reaction time was 30 min,and the substrate chromogenic time was 15 min.Through the detection of 11 kinds of samples,such as feed,it could be concluded that the semi-inhibitory concentration IC50 was less than 0.1μg/L,the recovery rate was 67%-116%,the sensitivity and effective detection were 0.03μg/L,the linear coefficient was greater than 0.99,and the in-board variation was less than 5%.The good test results showed that the improved method was feasible.