State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi 214122, China; International Joint Laboratory on Food Safety, Jiangnan University, Wuxi 214122, China.
J Hazard Mater. 2022 Jul 5;433:128750. doi: 10.1016/j.jhazmat.2022.128750. Epub 2022 Mar 23.
Deoxynivalenol (DON) is a typical mycotoxin in cereals and poses tremendous threats to the ecological environment and public health. Therefore, exploiting sensitive and robust analytical methods for DON is particularly important. Here, we fabricated a CRISPR-Cas12a-mediated luminescence resonance energy transfer (LRET) aptasensor to detect DON by using single-stranded DNA modified upconversion nanoparticles (ssDNA-UCNPs) as anti-interference luminescence labels and gold nanoparticle-decorated TiCT MXene nanosheets (MXene-Au) as enhanced quenchers. The DON aptamer can activate the trans-cleavage activity of Cas12a to indiscriminately cut nearby ssDNA-UCNPs into small fragments, which prevents ssDNA-UCNPs from adsorbing onto MXene-Au, and the upconversion luminescence (UCL) remains. Upon the binding of the aptamer with DON, the trans-cleavage activity of Cas12a was suppressed, and the ssDNA-UCNPs were not cleaved and easily adsorbed onto MXene-Au, which caused UCL quenching. Under optimized conditions, the limit of detection was determined to be 0.64 ng/mL with a linear range of 1 - 500 ng/mL. In addition, the sensor was successfully applied to detect DON in corn flour and Tai Lake water with recoveries of 96.2 - 105% and 95.2 - 104%, respectively. This platform achieves a sensitive and specific analysis of DON and greatly broadens the detection range of CRISPR-Cas sensors for non-nucleic acids hazards in the environment and food.
脱氧雪腐镰刀菌烯醇(DON)是谷物中的一种典型真菌毒素,对生态环境和公共健康构成了巨大威胁。因此,开发灵敏、稳健的 DON 分析方法尤为重要。在这里,我们构建了一种基于 CRISPR-Cas12a 的发光共振能量转移(LRET)适体传感器,通过使用经单链 DNA 修饰的上转换纳米粒子(ssDNA-UCNPs)作为抗干扰发光标记物和金纳米粒子修饰的 TiCT MXene 纳米片(MXene-Au)作为增强猝灭剂来检测 DON。DON 适体能激活 Cas12a 的转切割活性,从而将附近的 ssDNA-UCNPs 无差别地切割成小片段,阻止 ssDNA-UCNPs 吸附到 MXene-Au 上,从而保持上转换发光(UCL)不变。当适体与 DON 结合时,Cas12a 的转切割活性受到抑制,ssDNA-UCNPs 未被切割,容易吸附到 MXene-Au 上,导致 UCL 猝灭。在优化条件下,检测限为 0.64ng/mL,线性范围为 1-500ng/mL。此外,该传感器成功应用于玉米粉和太湖水中 DON 的检测,回收率分别为 96.2%-105%和 95.2%-104%。该平台实现了 DON 的灵敏和特异性分析,极大地拓宽了 CRISPR-Cas 传感器在环境和食品中非核酸危害物检测的范围。