Electrochemical biosensors combined with isothermal amplification for quantitative detection of nucleic acids

Miyuki Tabata, Bo Yao, Ayaka Seichi, Koji Suzuki, Yuji Miyahara

研究成果: Chapter

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In recent years, various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). The integration of isothermal amplification with electrical or electrochemical devices has enabled high-throughput nucleic acid-based assays with high sensitivity. We performed solid-phase rolling circle amplification (RCA) on the surface of a Au electrode, and detected RCA products in situ using chronocoulometry (CC) with [Ru (NH3)6]3+as the signaling molecule. Detection sensitivity for DNA and a microRNA (miR-143) was 100 fM and 1 pM, respectively. Furthermore, we conducted potentiometric DNA detection using an ethidium ion (Et+)-selective electrode (Et+ISE) for real-time monitoring of isothermal DNA amplification by primer-generation RCA (PG-RCA). The Et+ISE potential enabled real-time monitoring of the PG-RCA reaction in the range of 10 nM–1 μM of initial target DNA. Devices based on these electrochemical techniques represent a new strategy for replacing conventional PCR for on-site detection of nucleic acids of viruses or microorganisms.

元の言語English
ホスト出版物のタイトルMethods in Molecular Biology
出版者Humana Press Inc.
ページ135-151
ページ数17
DOI
出版物ステータスPublished - 2017

出版物シリーズ

名前Methods in Molecular Biology
1572
ISSN(印刷物)1064-3745

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Tabata, M., Yao, B., Seichi, A., Suzuki, K., & Miyahara, Y. (2017). Electrochemical biosensors combined with isothermal amplification for quantitative detection of nucleic acids. : Methods in Molecular Biology (pp. 135-151). (Methods in Molecular Biology; 巻数 1572). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6911-1_10