Label-free and Electrochemical Detection of Nucleic Acids Based on Isothermal Amplification in Combination with Solid-state pH Sensor☆
Label-free and Electrochemical Detection of Nucleic Acids Based on Isothermal Amplification in Combination with Solid-state pH Sensor☆
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DOI：
10.1016/j.proeng.2016.11.534
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
M. Tabata;Y. Katayama;F. Mannan;A. Seichi;Koji Suzuki;T. Goda;A. Matsumoto;Y. Miyahara
中科院分区：
文献类型：
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作者：
M. Tabata;Y. Katayama;F. Mannan;A. Seichi;Koji Suzuki;T. Goda;A. Matsumoto;Y. Miyahara
无标记核酸定量方法在疾病诊断或预后观察等临床应用中提供了潜在的工具，备受关注。在本研究中，我们将等温扩增法与无需标记的铱/氧化铱（Ir/IrOx）电极相结合，开发了一种更简单、性价比更高的核酸扩增电监测装置。制备的Ir/IrOx电极在不同的pH缓冲溶液下均表现出理想的Nernstian响应，且在带电蛋白存在下其电位响应受影响较小。此外，在恒定的反应温度下，通过检测扩增过程中释放的质子，我们成功地实时监测了靶DNA在0、10、100、1000 pM下的引物生成滚圈扩增（PG-RCA）。这种无标签和便携式设备可能会受到相当大的关注，作为一个有用的平台，在临床使用的点护理测试。
Label-free quantification methods for nucleic acids are attracting attention, since they provide potential tools in clinical applications such as disease diagnosis or prognosis observation. In this research, we developed a simpler and cost-effective electrical monitoring device of nucleic acid amplification, combining an isothermal amplification method and an Iridium/Iridium oxide (Ir/IrOx) electrode without labeling. The fabricated Ir/IrOx electrode showed ideal Nernstian response under the various pH buffer solutions, and its potential response was less affected even in the presence of charged proteins. Moreover, we successfully real-time monitored primer generation-rolling circle amplification (PG-RCA) at the 0, 10, 100, 1000 pM of target DNA by detection of released proton during amplification reaction at the constant reaction temperature. This label-free and portable device might be received considerable attention as a useful platform for a point-of-care testing in clinical use.