Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, Faculty of Medicine, University of Toronto, Toronto, Canada.
Anal Chem. 2013 Aug 6;85(15):7333-8. doi: 10.1021/ac401221f. Epub 2013 Jul 15.
Fast, sensitive nucleic acid sensors that enable direct detection of bacteria and diagnosis of infectious disease would offer significant advantages over existing approaches that employ enzymatic amplification of nucleic acids. We have developed chip-based microelectrodes that are highly effective for bacterial detection and have shown that they can capture and permit the analysis of large slow moving mRNA targets. Here, we explore new approaches to tune their analytical sensitivity and investigate the effect of sensor size, material composition, and probe density on the electrochemical signals obtained in the presence of bacteria. Sensor size can be varied from 10 to 100 μm, and this parameter can change detection limits obtained by a factor of 100. Changing the surface coating can also be used to tune sensitivity, with more nanostructured coatings yielding the most sensitive detectors. Moreover, we assessed performance of the sensors by tuning probe density. Varying the density of the immobilized probe had a dramatic effect on sensitivity, with sparse probe monolayers providing superior levels of performance. Overall, this study points to several factors that can be used to tune detection limits.
快速、灵敏的核酸传感器可以直接检测细菌并诊断传染病,这将比现有的核酸酶扩增方法具有显著优势。我们已经开发出基于芯片的微电极,它们在细菌检测方面非常有效,并已经证明它们可以捕获和允许对大型缓慢移动的 mRNA 靶标进行分析。在这里,我们探索了新的方法来调整它们的分析灵敏度,并研究了传感器尺寸、材料组成和探针密度对存在细菌时获得的电化学信号的影响。传感器的尺寸可以从 10 到 100μm 变化,这个参数可以将检测限改变 100 倍。改变表面涂层也可以用于调整灵敏度,具有更多纳米结构涂层的传感器具有最高的灵敏度。此外,我们通过调整探针密度来评估传感器的性能。固定探针的密度变化对灵敏度有显著影响,稀疏的探针单层提供了更好的性能水平。总的来说,这项研究指出了可以用于调整检测限的几个因素。