Real-Time Intracellular Analysis of Kanamycin Using Microaptasensors.
Real-Time Intracellular Analysis of Kanamycin Using Microaptasensors.
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DOI：
10.1021/acssensors.2c02427
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发表时间：
2023-02
期刊：
影响因子：
8.9
通讯作者：
V. Gupta;J. Dick
中科院分区：
文献类型：
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作者：
V. Gupta;J. Dick
随着耐多药细菌的出现，与感染相关的死亡人数正在上升。抗生素的过度使用及其泄漏到水道中已经将环境变成了水槽，导致细菌耐药性渗透到食物循环的各个层面。作为食物的主要来源之一，鱼和鱼产品如鱼卵必须研究其积累相关抗生素的能力。虽然先前已经研究了这些药物的积累，但是仍然需要在真实的时间中分析这些过程。基于电化学适体的传感器技术允许对小分子进行选择性的实时监测。在这里，我们报告的第一次使用的小型化电化学适配子为基础的传感器的被动吸收的氨基糖苷类抗生素，卡那霉素，在单个鲑鱼卵的分析。我们使用拉铂微电极，并通过树枝状金沉积增加电极尖端的表面积。与裸微电极相比，这些电极显示出在电极表面上的DNA固定增加了100倍。此外，与更广泛使用的宏传感器（r = 1 mm）相比，传感器在复杂生物介质中在延长的时间段内显示出改善的稳定性。传感器的范围被确定为从纳摩尔浓度延伸到鱼卵裂解物中卡那霉素的微摩尔浓度，并且当用于单个鲑鱼卵时，μ-适体传感器能够监测卡那霉素随时间的被动摄取。使用COMSOL Multiphysics软件模拟累积动力学。这项研究首次报道了使用电化学在单个细胞中实时被动吸收小分子的记录。
With the emergence of multidrug-resistant bacteria, infection-related death toll is on the rise. Overuse of antibiotics and their leakage into waterways have transformed the environment into a sink, resulting in bacterial resistance permeating through all tiers of the food cycle. As one of the primary sources of food, fish and fish products such as fish eggs must be studied for their ability to accumulate relevant antibiotics. While the accumulation of these pharmaceuticals has previously been studied, there remains a need to analyze these processes in real time. Electrochemical aptamer-based sensor technology allows for selective, real-time monitoring of small molecules. Herein, we report the first use of miniaturized electrochemical aptamer-based sensors for the analysis of the passive uptake of the aminoglycoside antibiotic, kanamycin, in single salmon eggs. We use pulled platinum microelectrodes and increase the surface area at the electrode tip through dendritic gold deposition. These electrodes showed a 100-fold increase in DNA immobilization on the electrode surface as compared to bare microelectrodes. Additionally, the sensors showed improved stability in complex biological media over an extended period of time when compared to the more widely used macrosensors (r = 1 mm). The sensor range was determined to extend from nanomolar to micromolar concentrations of kanamycin in fish egg lysate and when used in a single salmon egg the μ-aptasensors were able to monitor the passive uptake of kanamycin over time. The accumulation kinetics were simulated using COMSOL Multiphysics software. This research presents the first reported record of passive uptake of a small molecule in a single cell in real-time using electrochemistry.