同时检测AFB1和ZEA免疫层析试纸条的研制

采用柠檬酸三钠还原法制备胶体金,将其与抗AFB1和ZEA单克隆抗体进行偶联,并把偶联物、AFB1-STI、ZEA-OVA以及硝酸纤维素膜(NC膜)等组装成免疫层析试纸条,同时进行检测条件的最优化设计。胶体金偶联AFB1和ZEA抗体最适pH同为7.0,最适抗体浓度分别为5.0和2.5μg.mL-1,NC膜上包被AFB1-STI和ZEA-OVA的浓度分别1.0和2.0mg.mL-1,为该方法可同时定性检测样品中AFB1和ZEA,其最低检测限分别达到5μg.kg-1和50μg.kg-1;与黄曲霉毒素M1、脱氧血腐镰刀菌烯醇的交叉反应率小于1%。该试纸条具有可同时检测两种真菌毒素的特点,适用于同一样品中多种毒素的快速检测。

Development of immunochromatographic strip test for the rapid simultaneous detection of aflatoxin B1 and zearalenone

Aflatoxin B1（AFB1） and zearalenone（ZEA） monoclonal antibody（McAb） was conjugated to the colloidal gold particles as the detection reagent,respectively.Gold-McAb conjugate,AFB1-STI,ZEA-OVA and nitrocellulose membrane were assembled as immunochromatographic strip and optimize the test conditions.The optimal pH for both colloidal gold particles conjugated with AFB1 and ZEA McAb were 7.0 and the optimal protein concentration were 5.0 and 2.5 μg.mL-1,respectively.AFB1-STI and ZEA-OVA were coated on the nitrocellulose membrane and concentration were 1.0 and 2.0 mg.mL-1,respectively.This method can simultaneous qualitative detected AFB1 and ZEA in sample and the limit of detection were 5 μg.kg-1 and 50 μg.kg-1;There was no cross reaction to Aflatoxin M1 and deoxynivalenol.This strip is suitable for rapid detecting multiple toxins in one sample.