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利用基于石英晶体微天平的质量传感器实时监测核酸扩增反应。

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

机构信息

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

Institute for the Promotion of General Graduate Education (IPGE), Yamagata University, Yonezawa 992-8510, Japan.

出版信息

Biosensors (Basel). 2024 Mar 25;14(4):155. doi: 10.3390/bios14040155.

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.

摘要

核酸扩增反应,如聚合酶链反应(PCR),它使用 DNA 聚合酶来扩增单个双链 DNA 片段,是一种用于可视化特定基因组存在的有用技术。尽管荧光标记法主要用于 DNA 扩增,但应考虑其他检测方法,以进一步改进反应监测设备的微型化和降低成本。在这项研究中,石英晶体微天平(QCM)方法可测量纳克级质量,用于实时检测溶液中的核酸片段。这与基于重组酶聚合酶扩增(RPA)方法的等温核酸扩增反应相结合,允许在恒温下进行 DNA 扩增。当在带有固定化引物 DNA 链的 QCM 传感器板上启动 DNA 扩增反应时,与未固定化的引物 DNA 相比,观察到质量显著增加。结果表明,QCM 对扩增 DNA 片段的原位检测具有足够的灵敏度。将便携式 QCM 装置与 RPA 相结合,为检测核酸提供了一种灵敏的即时检测方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa6b/11048521/5efc3131b5f9/biosensors-14-00155-g001.jpg

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