Meng Jiajia, Li Ruijiao, Chen Xinyue, Wang Xinyi, Huang Qingwen, Nie Dongxia, Guo Dakai, Zhao Zhihui, Han Zheng
Institute for Agro-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1000 Jingqi Road, Shanghai 201403, China.
Institute for Agro-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1000 Jingqi Road, Shanghai 201403, China.
Food Chem. 2025 May 1;473:143042. doi: 10.1016/j.foodchem.2025.143042. Epub 2025 Jan 24.
Deoxynivalenol (DON) is one of the most harmful mycotoxins that poses great health threats to human and animals. Herein, a simple and sensitive magnetic beads-based fluorescent biosensor was successfully prepared for detection of DON in cereals. A stable double-stranded DNA (dsDNA, biotin-sDNA+FAM-cDNA/AP) was formed on the surface of streptavidin-coated magnetic beads (SMBs). DON could specifically bind to aptamer probe (AP), thereby releasing cDNA labeled with 6-carboxyfluorescein (FAM) (FAM-cDNA). After magnetic separation, the amount of FAM-cDNA was analyzed to achieve the detection of DON. Under the optimal conditions, the established aptasensor displayed excellent performance for DON analysis with the nice linear relationship (R = 0.9972) in the range of 0.1-50 ng/mL, high sensitivity (detection limit of 0.033 ng/mL) and selectivity. The total analysis time including samples preparation was around 200 min. The designed sensor was validated by determining the recoveries (95.48-112.48 %) in real paddy, wheat and maize samples, and its applicability to them was also verified by super-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The current study proposed a promising strategy for DON detection, and could easily be extended for the analysis of other targets through the rational design of the sequences of aptamer probe and cDNA.
脱氧雪腐镰刀菌烯醇（DON）是对人类和动物健康构成重大威胁的最有害霉菌毒素之一。在此，成功制备了一种基于磁珠的简单灵敏荧光生物传感器，用于检测谷物中的DON。在链霉亲和素包被的磁珠（SMB）表面形成了稳定的双链DNA（dsDNA，生物素-sDNA+FAM-cDNA/AP）。DON能特异性结合适配体探针（AP），从而释放出用6-羧基荧光素（FAM）标记的cDNA（FAM-cDNA）。经过磁分离后，分析FAM-cDNA的量以实现对DON的检测。在最佳条件下，所建立的适配体传感器对DON分析表现出优异的性能，在0.1-50 ng/mL范围内具有良好的线性关系（R = 0.9972）、高灵敏度（检测限为0.033 ng/mL）和选择性。包括样品制备在内的总分析时间约为200分钟。通过测定实际水稻、小麦和玉米样品中的回收率（95.48-112.48%）对所设计的传感器进行了验证，并用超高效液相色谱串联质谱法（UPLC-MS/MS）验证了其对这些样品的适用性。本研究提出了一种有前景的DON检测策略，并且通过合理设计适配体探针和cDNA的序列，可轻松扩展用于其他目标物的分析。
