Electrochemical Impedimetric Real-Time Polymerase Chain Reactions Using Anomalous Charge Transfer Enhancement
Electrochemical Impedimetric Real-Time Polymerase Chain Reactions Using Anomalous Charge Transfer Enhancement
复制标题
使用异常电荷转移增强的电化学阻抗实时聚合酶链反应
DOI：
10.1021/acs.analchem.2c01659
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发表时间：
2022
影响因子：
7.4
通讯作者：
Yamashita Ichiro
中科院分区：
文献类型：
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作者：
Nobusawa Kazuyuki;Han Huan-Wen;Takei Fumie;Chu Ting-Chieh;Hashida Noriyasu;Yamashita Ichiro
基于我们最近的发现，DNA 嵌入剂 [Ru(bpy)2DPPZ]2+ 异常增强氧化还原介体 K4[Fe(CN)6]/K3[Fe(CN)6] 和碳电极之间的电荷转移，我们开发了一种用于实时检测聚合酶链反应 (PCR) 的新电化学阻抗方法。将 3 mM [Fe(CN)6]3–/4– 和 5 μM [Ru(bpy)2DPPZ]2+ 添加到 PCR 溶液中，并在每次伸长热循环时进行电化学阻抗谱 (EIS) 测量。由于溶液中存在 [Ru(bpy)2DPPZ]2+，电荷转移电阻 (Rct) 最初较低。随着 PCR 的进行，扩增子 dsDNA 呈指数级产生，并且插入的 [Ru(bpy)2DPPZ]2+ 离子（可检测为陡峭的 Rct）在特定热循环下增加，具体取决于模板 DNA 的量。每个热循环的 Rct 增加量 ΔRct 在与光学检测相同的热循环中显示出峰值，证明可以通过阻抗测量来准确地实时监测 PCR。这种简单的方法将实现具有成本效益的便携式 PCR 设备。
We developed a new electrochemical impedimetric method for the real-time detection of polymerase chain reactions (PCR) based on our recent discovery that the DNA intercalator, [Ru(bpy)2DPPZ]2+, anomalously enhances charge transfer between redox mediators, K4[Fe(CN)6]/K3[Fe(CN)6], and a carbon electrode. Three mM [Fe(CN)6]3–/4–and 5 μM [Ru(bpy)2DPPZ]2+were added to the PCR solution, and electrochemical impedance spectroscopy (EIS) measurements were performed at each elongation heat cycle. The charge transfer resistance (Rct) was initially low due to the presence of [Ru(bpy)2DPPZ]2+in the solution. As PCR progressed, amplicon dsDNA was produced exponentially, and intercalated [Ru(bpy)2DPPZ]2+ions, which could be detected as a steep Rct, increased at specific heat cycles depending on the amount of template DNA. The Rct increase per heat cycle, ΔRct, showed a peak at the same heat cycle as optical detection, proving that PCR can be accurately monitored in real time by impedance measurement. This simple method will enable a cost-effective and portable PCR device.