Chapter title |
Electrochemical Biosensors Combined with Isothermal Amplification for Quantitative Detection of Nucleic Acids
|
---|---|
Chapter number | 10 |
Book title |
Biosensors and Biodetection
|
Published in |
Methods in molecular biology, March 2017
|
DOI | 10.1007/978-1-4939-6911-1_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6910-4, 978-1-4939-6911-1
|
Authors |
Miyuki Tabata, Bo Yao, Ayaka Seichi, Koji Suzuki, Yuji Miyahara |
Editors |
Ben Prickril, Avraham Rasooly |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 14 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 3 | 21% |
Researcher | 3 | 21% |
Unspecified | 2 | 14% |
Student > Bachelor | 1 | 7% |
Unknown | 5 | 36% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 3 | 21% |
Unspecified | 2 | 14% |
Chemistry | 2 | 14% |
Chemical Engineering | 1 | 7% |
Engineering | 1 | 7% |
Other | 0 | 0% |
Unknown | 5 | 36% |