A Thermus thermophilus argonaute-coupling exponential amplification assay for ultrarapid analysis of circulating tumor DNA

作者全名："Fang, Jie; Yuan, Changjing; Luo, Xizi; He, Zhe; Fu, Weiling"

作者地址："[Fang, Jie; Luo, Xizi; He, Zhe; Fu, Weiling] Third Mil Med Univ, Army Med Univ, Southwest Hosp, Dept Lab Med, Chongqing 400038, Peoples R China; [Fang, Jie] Chongqing Med Univ, Affiliated Hosp 2, Dept Lab Med, Chongqing 400010, Peoples R China; [Yuan, Changjing] Chongqing Med Univ, Affiliated Hosp 1, Dept Lab Med, Chongqing 400016, Peoples R China"

通信作者："Fu, WL (通讯作者)，Third Mil Med Univ, Army Med Univ, Southwest Hosp, Dept Lab Med, Chongqing 400038, Peoples R China.; Yuan, CJ (通讯作者)，Chongqing Med Univ, Affiliated Hosp 1, Dept Lab Med, Chongqing 400016, Peoples R China."

来源：TALANTA

ESI学科分类：CHEMISTRY

WOS号：WOS:001064425500001

JCR分区：Q1

影响因子：6.1

年份：2024

卷号：266

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：Thermus thermophilus argonaute; Exponential amplification reaction; Circulating tumor DNA; Terahertz spectroscopy; Lateral flow detection; Biosensor

摘要："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-flowbased 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."

基金机构：National Natural Science Foundation of China [81920108024]; Military logistics scientific research project [AWS17J010]; Project for frontline medical workers in epidemic prevention [2020FYYX022]

基金资助正文："This study was supported by National Natural Science Foundation of China (81920108024), the Military logistics scientific research project (AWS17J010) and Project for frontline medical workers in epidemic prevention (2020FYYX022)."