The simultaneous analysis of the levels of multiple microRNAs (miRNAs) is critical to the early diagnosis of cancer. However, this analysis is challenging because of the low concentrations of miRNAs and their high sequence homology. Here, we report a general and programmable diagnostic strategy for miRNA analysis: Thermus thermophilus Argonaute (TtAgo)-assisted exponential isothermal amplification for multiplex detection (TEAM). This system combines exponential isothermal amplification (EXPAR), for target amplification, with programmable TtAgo cleavage, for the generation of the reporting signal. The TEAM assay achieved attomolar sensitivity with a rapid turnaround time (30–35 min). Because of the single-nucleotide precision of TtAgo, the system demonstrated robust multiplex capability in the simultaneous detection of four miRNA targets and the classification of let-7 family members. The TEAM assay was superior in differentiating colorectal cancer patients from healthy individuals relative to the conventional EXPAR and reverse transcription polymerase chain reaction (RT-PCR) methods. This tunable and scalable approach is a powerful nucleic acid analysis tool that holds promise in scientific and clinical applications.

中文翻译：

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嗜热栖热菌 Argonaute 辅助指数等温扩增多重检测 (TEAM) 对 MicroRNA 的可编程分析

同时分析多个 microRNA (miRNA) 的水平对于癌症的早期诊断至关重要。然而，由于 miRNA 浓度低且序列同源性高，这种分析具有挑战性。在这里，我们报告了一种用于 miRNA 分析的通用且可编程的诊断策略：Thermus thermophilusArgonaute (TtAgo) 辅助指数等温扩增用于多重检测 (TEAM)。该系统结合了指数等温扩增 (EXPAR)，用于目标扩增，与可编程的 TtAgo 切割相结合，用于生成报告信号。TEAM 测定以快速周转时间（30-35 分钟）实现了阿托摩尔灵敏度。由于 TtAgo 的单核苷酸精度，该系统在同时检测四个 miRNA 靶标和分类 let-7 家族成员方面表现出强大的多重能力。相对于传统的 EXPAR 和逆转录聚合酶链反应 (RT-PCR) 方法，TEAM 检测在区分结直肠癌患者和健康个体方面具有优势。