谷物中伏马菌素B1残留的直接竞争ELISA检测方法研究

摘要

以抗伏马菌素B1(Fumonisin B1,FB1)单抗作为包被抗体,辣根过氧化物酶(HRP-FB1)作为标记抗原,建立检测谷物中FB1的直接竞争酶联免疫吸附法(dc-ELISA)；并与国家标准方法(HPLC,GB/T 255228-2010)进行比较,选取真菌毒素呕吐毒素、玉米赤霉烯酮、黄曲霉毒素B1考察方法的特异性.结果显示:该dc-ELISA方法线性范围在200～800 ng/mL之间,回归方程为Y=-0.000 8X+0.978 9,决定系数R2为0.997 4,检出限为182.4 ng/mL,6份样品的批内和批间变异系数分别为7.4％和7.7％,平均加标回收率为98.5％(n=6).样品检测显示dc-ELISA与HPLC方法具有良好的相关性(r=0.999 6),与其他真菌毒素无交叉反应.因此,dc-ELISA法可应用检测玉米等谷物中FB1的含量,适合大批量筛选样品.%Taking fumonisin B1 (FB1) monoclonal antibodies as the coating antibody and horse radish peroxidase as labeled antigen,we established a direct competitive ELISA(dc-ELISA) method for determining FB1 in grains,then compared the dc-ELISA method with the HPLC method,and studied the specificity of the method by using deoxynivalenol,aflatoxin B1 and zearalenone.The results showed that:the linear range of ELISA method was from 200 to 800 ng/mL; the regression equation was Y=0.000 8X+0.978 9,the determination coefficient R2 was 0.997 4; the detection limit was 182.4 ng/mL; the coefficients of variation of intra-assay and inter-assay of six samples were respectively 7.4％ and 7.7％; and the average recovery rate was 98.5％.The results showed that dc-ELISA method had good correlation with HPLC method (r=0.999 6),and had no cross-activity with other mycotoxins.Accordingly,the dc-ELISA method could be used for determining FB1 content in grains,such as maize,and was suitable for mass sample screening.