Qingdao Key Laboratory of Food Biotechnology, College of Food Science and Engineering, Ocean University of China, Qingdao 266404, PR China; Key Laboratory of Biological Processing of Aquatic Products, China National Light Industry, Qingdao 266404, PR China.
Qingdao Key Laboratory of Food Biotechnology, College of Food Science and Engineering, Ocean University of China, Qingdao 266404, PR China; Key Laboratory of Biological Processing of Aquatic Products, China National Light Industry, Qingdao 266404, PR China.
J Hazard Mater. 2023 Sep 15;458:131995. doi: 10.1016/j.jhazmat.2023.131995. Epub 2023 Jul 4.
Developing simultaneous detection methods for multiple targets is crucial for the field of food analysis. Herein, enrofloxacin (ENR) and ciprofloxacin (CIP) were taken as model targets. For the first time, a strategy to generate group-specific split aptamers was established by revealing and splitting the critical binding domain, and the split aptamers were exploited to design a four-way DNA junction (4WJ) which could regulate the enzymatic activity of chitosan oligosaccharide (COS)-AuNPs nanozyme to develop a colorimetric aptasensor. A pair of split aptamers were obtained for ENR (K = 15.00 nM) and CIP (K = 4.870 nM). The mechanism of COS binding with double-stranded DNA in the 4WJ was elucidated. Under optimal conditions, the colorimetric aptasensor enabled a wide linear detection range of 1.4-1400 nM and a limit of detection (LOD) of 321.1 pM and 961.0 pM towards ENR and CIP, respectively, which exhibited excellent sensitivity, selectivity, and availability in detecting ENR/CIP in seafood. This study expands the general strategies for generating robust aptamers and nanozyme complex and provides a good reference for developing multi-target detection methods.
开发用于多个目标物的同时检测方法对于食品分析领域至关重要。在此，以恩诺沙星（ENR）和环丙沙星（CIP）为模型目标物。首次通过揭示和拆分关键结合域建立了一种产生组特异性分割适体的策略，并利用分割适体设计了一种四向 DNA 连接体（4WJ），该连接体可调节壳聚糖寡糖（COS）-AuNPs 纳米酶的酶活性，以开发比色适体传感器。获得了一对针对 ENR（K = 15.00 nM）和 CIP（K = 4.870 nM）的分割适体。阐明了 4WJ 中 COS 与双链 DNA 结合的机制。在最佳条件下，比色适体传感器对 ENR 和 CIP 的线性检测范围分别为 1.4-1400 nM 和检测限（LOD）分别为 321.1 pM 和 961.0 pM，在检测海鲜中的 ENR/CIP 时表现出优异的灵敏度、选择性和实用性。该研究扩展了生成稳健适体和纳米酶复合物的一般策略，并为开发多目标物检测方法提供了良好的参考。
