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Article

Real-Time Monitoring of a Nucleic Acid Amplification Reaction Using a Mass Sensor Based on a Quartz-Crystal Microbalance

by
Hideto Kumagai
1 and
Hiroyuki Furusawa
1,2,*
1
Graduate School of Organic Materials Science, Yamagata University, Yonezawa 992-8510, Japan
2
Institute for the Promotion of General Graduate Education (IPGE), Yamagata University, Yonezawa 992-8510, Japan
*
Author to whom correspondence should be addressed.
Biosensors 2024, 14(4), 155; https://doi.org/10.3390/bios14040155
Submission received: 29 January 2024 / Revised: 21 March 2024 / Accepted: 23 March 2024 / Published: 25 March 2024
(This article belongs to the Special Issue Development of Novel Biosensors for Point-of-Care Detection)

Abstract

Nucleic acid amplification reactions such as polymerase chain reaction (PCR), which uses a DNA polymerase to amplify individual double-stranded DNA fragments, are a useful technique for visualizing the presence of specific genomes. Although the fluorescent labeling method is mainly used with DNA amplification, other detection methods should be considered for further improvements, such as miniaturization and cost reduction, of reaction-monitoring devices. In this study, the quartz-crystal microbalance (QCM) method, which can measure nanogram-order masses, was applied for the real-time detection of DNA fragments in a solution with nucleic acids. This was combined with an isothermal nucleic acid amplification reaction based on the recombinase polymerase amplification (RPA) method, which allowed DNA amplification at a constant temperature. When the DNA amplification reaction was initiated on a QCM sensor plate with an immobilized primer DNA strand, a significant increase in mass was observed compared to when the primer DNA was not immobilized. QCM was shown to be sufficiently sensitive for the in situ detection of amplified DNA fragments. Combining a portable QCM device and RPA offers a sensitive point-of-care method for detecting nucleic acids.
Keywords: polymerase chain reaction; quartz-crystal microbalance; mass sensor; isothermal nucleic acid amplification reaction; recombinase polymerase amplification; DNA fragment detection polymerase chain reaction; quartz-crystal microbalance; mass sensor; isothermal nucleic acid amplification reaction; recombinase polymerase amplification; DNA fragment detection

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MDPI and ACS Style

Kumagai, H.; Furusawa, H. Real-Time Monitoring of a Nucleic Acid Amplification Reaction Using a Mass Sensor Based on a Quartz-Crystal Microbalance. Biosensors 2024, 14, 155. https://doi.org/10.3390/bios14040155

AMA Style

Kumagai H, Furusawa H. Real-Time Monitoring of a Nucleic Acid Amplification Reaction Using a Mass Sensor Based on a Quartz-Crystal Microbalance. Biosensors. 2024; 14(4):155. https://doi.org/10.3390/bios14040155

Chicago/Turabian Style

Kumagai, Hideto, and Hiroyuki Furusawa. 2024. "Real-Time Monitoring of a Nucleic Acid Amplification Reaction Using a Mass Sensor Based on a Quartz-Crystal Microbalance" Biosensors 14, no. 4: 155. https://doi.org/10.3390/bios14040155

APA Style

Kumagai, H., & Furusawa, H. (2024). Real-Time Monitoring of a Nucleic Acid Amplification Reaction Using a Mass Sensor Based on a Quartz-Crystal Microbalance. Biosensors, 14(4), 155. https://doi.org/10.3390/bios14040155

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