玉米中伏马毒素B1、B2间接竞争酶联免疫吸附方法的建立

An indirect competitive ELISA method for detecting fumonisin B1 and B2 in maize

基于伏马毒素B1(fumonisin B1,FB1)单克隆抗体建立了间接竞争酶联免疫吸附法(indirect competitive enzyme linked immunosorbent assay,ic-ELISA),用于玉米中伏马毒素的初步筛查。该方法优化了抗原包被浓度和单抗体稀释倍数、抗原包被条件、酶标二抗稀释倍数、底物作用时间。结果显示,线性检测范围为1.41~54.2ng/m L,最低检测限为24.5μg/kg,实测玉米样品加标回收率为89.36%~108.54%。交叉反应结果显示,与伏马毒素B2(fumonisin B2,FB2)交叉反应率为129.88%,与其他真菌毒素交叉反应均小于10%,该ic-ELISA方法可对玉米样品中的FB1与FB2总量进行初筛。

Based on a monoclonal antibody( McAb),an indirect competitive enzyme linked immunosorbent assay( ic-ELISA) was developed for detection of fumonisin B1( FB1) and B2( FB2).Experimental parameters,including amount of the coating antigen and McAb,coating conditions of the antigen,dilution rate of horseradish labeled secondary antibody( HRP-IgG) and time of chromogenic reaction were optimized. The linear detection range of the ic-ELISA was 1. 41-54. 2 ng/m L. The detection limit was 24. 5 μg/kg,and the recovery rate of FB1 and FB2 from maize samples was from89. 36% to 108. 54%. The cross-reaction rate of this ic-ELISA with FB1 and FB2 were 100% and129. 88% respectively,and lower than 10% with other mycotoxins. The ic-ELISA can be used for detection of the total amount of FB1 and FB2 in corn samples.