基于蛋白金属编码的适体夹心法对多真菌毒素的快速电化学分析研究

The consumption of mycotoxins-contaminated diet is potentially a health hazards for both humans and animals through induction of acute and chronic effects that have teratogenic, carcinogenic or immunosuppressive impact. Among mycotoxins of gtretest public health and agroeconomic significance are aflatoxins B1(AFB1) and ochratoxin A(OTA). Research on easy to use, sensitive, rapid and automated detection technology for mycotoxins is of great practical significance to monitor the mycotoxins and strengthen the food safety. This investigation is carried out for rapid and simutaneous determination of the AFB1 and OTA levels in food using graphene-Au grafted recombinant apoferritin-encoded metallic labels as signal tags and dual-aptemer magnetic beads as capture probes. We intend to focus on enhancing the capture efficiency and the resulting sensitivity of the proposed assay, by taking the magnetic beads(MBs) as the solid phase and splitting the aptamer into two nucleic acid sequence to construct the sandwich system. Finally, the proposed assay will be further evaluated through collaborative detection of the same practical sample using the official analysis method of HPLC or HPLC-MS. Hopefully, the project may provide new ideas and help pave the way for the R&D of Chinese detection technology and instrument for mycotoxins with independent intellectual property.

真菌毒素对于人体和动物都具有致癌、致畸、致突变等毒害作用，其中黄曲霉毒素B1（AFB1）与赭曲霉毒素A(OTA)的毒性最大。研究简便、灵敏、快速的真菌毒素自动化检测技术对于监控真菌毒素、加强食品安全保障有着重大的现实意义。本项目提出以石墨烯-纳米金复合物修饰的去铁铁蛋白金属分子作为标记物，结合修饰有适配体的磁珠作为探针的电化学夹心体系用于AFB1与OTA的同时检测。拟采用磁珠作为固相载体，核酸适配体进行剪切分为两段作为夹心体系的两组核酸片段，重点提高小分子的捕获效率和检测灵敏度。最后同步检测农贸市场抽检的实际样品，并与官方标准方法比较结果，力求为我国研制足有自主知识产权的真菌毒素检测技术和仪器提供新思路。

真菌毒素对于人体和动物都具有致癌、致畸、致突变等毒害作用，其中黄曲霉毒素B1（AFB1）的毒性最大。研究简便、灵敏、快速的真菌毒素自动化检测技术对于监控真菌毒素、加强食品安全保障有着重大的现实意义。本项目提出以石墨烯-纳米金复合物修饰的去铁铁蛋白金属分子作为标记物，结合修饰有适配体的磁珠作为探针的电化学夹心体系用于AFB1的检测。采用一步合成法制备了纳米金石墨烯修饰电极并据此构建了无需汞/铋离子快速检测实际样本中AFB1的方法。在1–90 μg/L的范围内，溶出电流与铅离子浓度之间呈现良好的线性关系，检测限为0.168 μg/L。使用此电极对实际样本分别进行铅离子加标回收率实验，水样中回收率为93.75%-109.73%，土壤样本的回收率为93.82%-109.9%。