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 language | English |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 135-151 |
| Number of pages | 17 |
| Volume | 1572 |
| DOIs | |
| Publication status | Published - 2017 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1572 |
| 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
Electrochemical biosensors combined with isothermal amplification for quantitative detection of nucleic acids. / Tabata, Miyuki; Yao, Bo; Seichi, Ayaka; Suzuki, Koji; Miyahara, Yuji.
Methods in Molecular Biology. Vol. 1572 Humana Press Inc., 2017. p. 135-151 (Methods in Molecular Biology; Vol. 1572).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Electrochemical biosensors combined with isothermal amplification for quantitative detection of nucleic acids
AU - Tabata, Miyuki
AU - Yao, Bo
AU - Seichi, Ayaka
AU - Suzuki, Koji
AU - Miyahara, Yuji
PY - 2017
Y1 - 2017
N2 - 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.
AB - 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.
KW - Biosensor
KW - Chronocoulometry
KW - Ion selective electrode
KW - Isothermal nucleic acid amplification
KW - MicroRNA
KW - PCR
UR - http://www.scopus.com/inward/record.url?scp=85015674175&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85015674175&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-6911-1_10
DO - 10.1007/978-1-4939-6911-1_10
M3 - Chapter
AN - SCOPUS:85015674175
VL - 1572
T3 - Methods in Molecular Biology
SP - 135
EP - 151
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -