同时检测伏马菌素B1和呕吐毒素的通用探针免疫层析卡研究

Study on general probe immunochromatographic assay strip for simultaneous detection of fumonisin B1 and deoxynivalenol

中文关键词:  真菌毒素  多残留检测  免疫层析检测  通用探针  双功能抗原

英文关键词:mycotoxins  multi-residual detection  immunochromatography assay  general probe  bifunctional antigen

中文摘要:

      目的 研发同时检测伏马菌素B1 (fumonisin B1, FB1)和呕吐毒素(deoxynivalenol, DON)的通用探针免疫层析卡。方法 通过标记物修饰在融合真菌毒素模拟表位的甘露糖结合蛋白(mannose binding protein, MBP)制备双功能抗原, 引导针对标记物抗体的量子点微球通用探针, 与检测线上的毒素抗体形成“夹心免疫复合物”, 从而产生检测信号。结果 标记物对MBP的反应摩尔比为3.0:1.0, 所制备的2种双功能抗原具有最优的双抗体结合活性。“毒素抗体/双功能抗原/通用探针”免疫复合物在2条检测线(T线)都形成荧光信号; 只需调整2种毒素抗体的点阵浓度, FB1和DON的T线完全抑制的阈值统一优化为预定的5.0 ng/mL。结论 基于双功能抗原引导的通用探针的免疫层析检测, 避免了分别制备单一特异性探针的烦琐, 各T线检测阈值优化过程简便, 适合真菌毒素多残留检测免疫层析试剂的标准化制备。

英文摘要:

      Objective To develop a general probe immunochromatographic assay strip for the simultaneous detection of fumonisin B1 (FB1) and deoxynivalenol (DON). Methods A chemical maker was modified to mannose binding protein (MBP) fused with mycotoxin mimic epitope for the preparation of bifunctional antigen, antibodies with specificity to the maker were labeled to the quantum dot beans to obtain general probes, the bifunctional antigens induced general probes to form a ‘sandwich immune complex’ with mycotoxin antibodies for generation of fluorescent signal on the testing line. Results The optimized reacting mole ratio of maker to MBP for bifunctional antigen was 3.0:1.0, "Toxin antibody/bifunctional antigen/general probe" immune complex forms fluorescent signals on both testing lines (T-line); the detection limits of FB1 and DON were optimized to the preset concentration of 5.0 ng/mL respectively only by adjusting the antibody levels. Conclusion The immunochromatographic assay based on the general probe coupled with bifunctional testing antigens avoids the complexity of preparing by individual specific probe for each mycotoxin, respectively. The optimization of the threshold for each T-line was simple. Thus, this method is suitable for the standardized development of immunochromatographic assays for multi-residual detection of mycotoxins with compound contamination.