Department of Chemistry, Fudan University, 200438 Shanghai, China.
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100101 Beijing, China.
Anal Chem. 2023 Sep 12;95(36):13401-13406. doi: 10.1021/acs.analchem.3c01990. Epub 2023 Aug 11.
Assays for the molecular detection of miRNAs are typically constrained by the level of multiplexing, especially in a single tube. Here, we report a general and programmable diagnostic platform by combining mesophilic Argonaute (CpAgo) with exponential isothermal amplification (EXPAR), which is a dual-signal amplification strategy, allowing for the rapid and sensitive detection of multiple miRNAs with single-nucleotide discrimination in one pot. The pAgo-based ne-ot (COP) assay achieved a limit of detection of 1 zM miRNA within 30 min of turnaround time and a wide concentration range. This COP assay was applied to simultaneously detect four miRNAs in a single tube from clinical serum samples, showing superior analytical performance in distinguishing colorectal cancer patients from healthy individuals. This programmable, one-pot, multiplex, rapid, and specific strategy offers great promise in scientific research and clinical applications.
用于分子检测 miRNA 的分析通常受到多重检测的限制，尤其是在单个管中。在这里，我们报告了一种通用的可编程诊断平台，该平台通过将嗜温 Argonaute（CpAgo）与指数等温扩增（EXPAR）相结合，实现了快速和敏感的检测，可在一个管中对多种 miRNA 进行单核苷酸区分。基于 pAgo 的 ne-ot（COP）分析在 30 分钟的检测时间内实现了 1 zM miRNA 的检测限，且具有较宽的浓度范围。该 COP 分析应用于从临床血清样本中同时检测四种 miRNA，在区分结直肠癌患者和健康个体方面表现出优异的分析性能。这种可编程、一锅法、多重、快速和特异的策略在科学研究和临床应用中具有广阔的前景。
