Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, Universitätsstraße 31, 93053 Regensburg, Germany.
Department of Chemistry, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Lab Chip. 2022 Sep 27;22(19):3721-3733. doi: 10.1039/d2lc00593j.
High performance, laser-induced graphene (LIG) electrodes were integrated into adhesive tape-based microfluidic channels to realize both electrochemical (EC) and electrochemiluminescent (ECL) detection approaches. This provides strategies for low limits of detection, simple hardware requirements and inexpensive fabrication, which are characteristics required for assays in the competitive point-of-care (POC) sensor field. Here, electrode design and microchannel dimensions were studied and a DNA hybridization assay with liposomes for signal amplification was developed for the specific detection of DNA derived from as the model analyte. Liposomes entrapped either Ru(bpy) or K[Fe(CN)] generating ECL- and EC-signal amplification, respectively. This new microchip provided all desirable analytical figures of merit needed for POC applications. Specifically, a desirable one-step assay was designed which provided a limit of detection of 3 pmol L for the ECL and 47 pmol L for the EC approach and furthermore enabled highly specific detection considering that at room temperature in this simple setup a single nucleotide polymorphism resulted in a signal decrease of 58%, whereas a decrease of > 98% was observed for non-matching sequences present in 10-fold excess. Direct detection in various matrices ranging from drinking water to soil extracts was also achieved. It is concluded that the simple and inexpensive fabrication in combination with signal amplification strategies makes these concepts relevant for on-site pathogen detection in resource-limited environments.
高性能激光诱导石墨烯 (LIG) 电极集成到基于胶带的微流道中,以实现电化学 (EC) 和电化学发光 (ECL) 检测方法。这为低检测限、简单的硬件要求和廉价的制造提供了策略,这些是竞争激烈的即时检测 (POC) 传感器领域分析物所需的特性。在这里,研究了电极设计和微通道尺寸,并开发了一种带有脂质体用于信号放大的 DNA 杂交分析物,用于特异性检测作为模型分析物的 DNA。脂质体包封了 Ru(bpy) 或 K[Fe(CN)],分别产生 ECL 和 EC 信号放大。这种新的微芯片提供了适用于 POC 应用的所有理想分析性能。具体来说,设计了一种理想的一步法测定法,该法为 ECL 提供了 3 pmol L 的检测限,为 EC 方法提供了 47 pmol L 的检测限,并且考虑到在这种简单设置下,室温下单个核苷酸多态性导致信号降低 58%,而对于存在 10 倍过量的非匹配序列,则观察到信号降低>98%。还实现了直接在各种基质(从饮用水到土壤提取物)中的检测。结论是,简单且廉价的制造与信号放大策略相结合,使得这些概念在资源有限的环境中用于现场病原体检测具有相关性。