分子检测miRNA的测定方法通常受到多重复线的限制，尤其是在一个试管内。在这里，我们报告了一种通用的、可编程的诊断平台，该平台通过组合中温梭状芽孢杆菌 Argonaute（CpAgo）与指数等温扩增（EXPAR）来实现，这是一种双信号放大策略，可以在一个容器中迅速、灵敏地检测多个miRNA，并实现单核苷酸鉴定。基于CpAgo的一锅式（COP）测定法在30分钟内实现了1 zM miRNA的检测限度，并具有广泛的浓度范围。该COP测定法应用于从临床血清样品中同时检测四个miRNA，在区分结直肠癌患者和健康人群方面表现出优越的分析性能。这种可编程、一锅式、多重复、快速和特异性策略在科学研究和临床应用中具有巨大潜力。

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 Clostridium perfringens 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 CpAgo-based One-Pot (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.