Wang Chen, Liu Jingliang, Kong Jinming, Zhang Xueji
School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing, 210094, PR China.
School of Environmental Science, Nanjing XiaoZhuang University, Nanjing, 211171, PR China.
Anal Chim Acta. 2020 Nov 1;1136:19-24. doi: 10.1016/j.aca.2020.08.035. Epub 2020 Aug 28.
In this work, a novel electrochemical biosensor based on nitronyl nitroxide monoradical 2,2,6,6-tetramethylpiperidine 1-Oxyl (TEMPO) as new electrochemical label for facile nucleic acids detection is developed. This fast and convenient functional microelectrode was designed by fixing the capture probe peptide nucleic acid (PNA) and using the coordination interaction of Zr with both phosphate groups and carboxyl groups. Differential pulse voltammetry (DPV) was used to study the oxidation current of TEMPO which was combined with the electrode surface and labeled. TEMPO electrochemical signal related to target deoxyribonucleic acid (tDNA) concentration was finally detected when tDNA was added on the surface of glassy carbon electrode (GCE). The detection principle, optimization of key factors and performance analysis of the biosensor are also discussed. A great linear relation is acquired within the scope of 10 pM-100 nM under optimal conditions and the detection limit of this experiment is calculated as low as 2.57 pM (R = 0.996). In addition, complex serum samples were used to explore the practical application of this experiment. The results show the developed electrochemical DNA biosensor has wide application prospects in nucleic acids detection and clinical analysis.
在这项工作中,开发了一种基于硝酰氮氧自由基单自由基2,2,6,6 - 四甲基哌啶1 - 氧基(TEMPO)作为新型电化学标记物的新型电化学生物传感器,用于便捷的核酸检测。这种快速便捷的功能微电极是通过固定捕获探针肽核酸(PNA)并利用Zr与磷酸基团和羧基的配位相互作用设计而成。采用差分脉冲伏安法(DPV)研究与电极表面结合并标记的TEMPO的氧化电流。当在玻碳电极(GCE)表面加入目标脱氧核糖核酸(tDNA)时,最终检测到与目标脱氧核糖核酸(tDNA)浓度相关的TEMPO电化学信号。还讨论了生物传感器的检测原理、关键因素的优化及性能分析。在最佳条件下,在10 pM - 100 nM范围内获得了良好的线性关系,本实验的检测限计算低至2.57 pM(R = 0.996)。此外,使用复杂血清样本探索了本实验的实际应用。结果表明,所开发的电化学DNA生物传感器在核酸检测和临床分析中具有广阔的应用前景。