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

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

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages135-151
Number of pages17
Volume1572
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1572
ISSN (Print)10643745

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Keywords

  • Biosensor
  • Chronocoulometry
  • Ion selective electrode
  • Isothermal nucleic acid amplification
  • MicroRNA
  • PCR

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

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