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

Hideto Kumagai1, Hiroyuki Furusawa1,2

  • 1Graduate School of Organic Materials Science, Yamagata University, Yonezawa 992-8510, Japan.

Biosensors
|April 26, 2024
PubMed
Summary

Quartz-crystal microbalance (QCM) combined with recombinase polymerase amplification (RPA) enables real-time DNA detection. This sensitive method offers a cost-effective, point-of-care solution for nucleic acid analysis.

Keywords:
DNA fragment detectionisothermal nucleic acid amplification reactionmass sensorpolymerase chain reactionquartz-crystal microbalancerecombinase polymerase amplification

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Nucleic acid amplification, like polymerase chain reaction (PCR), is vital for detecting specific genomes.
  • Current methods often rely on fluorescent labeling, limiting miniaturization and cost reduction.
  • Alternative detection strategies are needed for improved reaction-monitoring devices.

Purpose of the Study:

  • To apply the quartz-crystal microbalance (QCM) method for real-time DNA fragment detection.
  • To integrate QCM with isothermal nucleic acid amplification using recombinase polymerase amplification (RPA).
  • To assess the sensitivity and applicability of QCM-RPA for in situ nucleic acid detection.

Main Methods:

  • Utilized quartz-crystal microbalance (QCM) for mass-based detection of nanogram-order quantities.
  • Employed recombinase polymerase amplification (RPA) for isothermal DNA amplification at a constant temperature.
  • Immobilized primer DNA strands on a QCM sensor plate to initiate amplification.

Main Results:

  • Observed a significant mass increase on the QCM sensor when DNA amplification was initiated with immobilized primers.
  • Demonstrated that QCM is sufficiently sensitive for in situ detection of amplified DNA fragments.
  • Validated the combined QCM and RPA approach for real-time nucleic acid monitoring.

Conclusions:

  • The QCM method provides sensitive, real-time detection of DNA amplification.
  • Combining portable QCM devices with RPA offers a promising point-of-care nucleic acid detection method.
  • This integrated approach facilitates miniaturization and cost reduction in diagnostic devices.