循环肿瘤DNA（ctDNA）是一种非侵入性液体活检生物标志物，具有重要的临床和生物学信息，但现有的检测技术费用昂贵、复杂且耗时。本文报道了一种超快速、敏感且简单的方法，我们将其称为Thermus thermophilus argonaute耦合指数增殖反应（TtAgo-CEAR），可选择性地扩增突变的ctDNA。针对七种Kirsten Rat Sarcoma-2病毒（KRAS）点突变，本策略可以在仅需16分钟、无需前期PCR扩增的情况下，实现以aM级敏感性和单核苷酸特异性的简易检测。我们还证明了TtAgo耦合分析可轻松用于基于太赫兹光谱和侧流法的读出。我们展示了通过小鼠模型检测到的ctDNA浓度可以响应血清样品中疾病负担的变化。可以预见，这种TtAgo-CEAR方法对于快速诊断和监测各种恶性肿瘤具有巨大潜力。版权所有© 2023 Elsevier B.V.发表。

Circulating tumor DNA (ctDNA) is a noninvasive biomarker for liquid biopsy with important clinical and biological information, but existing detection techniques are expensive, complex and quite time-consuming. Here, we report an ultrarapid, sensitive and simple method, which we term Thermus thermophilus argonaute-coupling exponential amplification reaction (TtAgo-CEAR), that selectively amplifies mutated ctDNA. Aiming at seven Kirsten rat sarcoma-2 virus (KRAS) point mutations, the present strategy allows for easy detection with attomolar sensitivity and single-nucleotide specificity within as little as 16 min without prior PCR amplification. We also demonstrate that TtAgo-coupling assay is easily adaptable to Terahertz spectroscopy-based and lateral-flow-based readout. We show that the detected ctDNA concentrations by mouse models can respond to the variations of disease burden in serum samples. It is envisioned that this TtAgo-CEAR approach has great potential for rapid diagnosis and monitoring of diverse malignant tumors.Copyright © 2023. Published by Elsevier B.V.