对液体活检中细胞外囊泡 (EV) 中发现的信使 RNA (mRNA) 进行测序可以提供体细胞突变、耐药谱和肿瘤复发等临床信息。尽管如此，由于液体活检中的丰度较低，EV mRNA 仍未得到充分利用，并且需要大量样本或专门的分析技术。在这里，我们介绍自扩增和 CRISPR 辅助操作来分析 EV（SCOPE），这是一个用于 EV mRNA 检测的平台。 SCOPE 利用 Cas13 对目标 RNA 进行 CRISPR 介导的识别来启动复制和信号放大，在保持单核苷酸分辨率的同时实现亚阿摩尔检测限。作为概念验证，我们设计了针对 KRAS、BRAF、EGFR 和 IDH1 基因关键突变的探针，优化了单罐检测方案，并实施了用于多样本检测的自动化设备。我们验证了 SCOPE 在动物模型中检测早期肺癌的能力，监测结直肠癌患者的肿瘤突变负荷以及对胶质母细胞瘤患者进行分层。 SCOPE 可以加快读数，增强 EV 在精准肿瘤学中的临床应用。© 2024。作者获得 Springer Nature America, Inc. 的独家许可。

Sequencing of messenger RNA (mRNA) found in extracellular vesicles (EVs) in liquid biopsies can provide clinical information such as somatic mutations, resistance profiles and tumor recurrence. Despite this, EV mRNA remains underused due to its low abundance in liquid biopsies, and large sample volumes or specialized techniques for analysis are required. Here we introduce Self-amplified and CRISPR-aided Operation to Profile EVs (SCOPE), a platform for EV mRNA detection. SCOPE leverages CRISPR-mediated recognition of target RNA using Cas13 to initiate replication and signal amplification, achieving a sub-attomolar detection limit while maintaining single-nucleotide resolution. As a proof of concept, we designed probes for key mutations in KRAS, BRAF, EGFR and IDH1 genes, optimized protocols for single-pot assays and implemented an automated device for multi-sample detection. We validated SCOPE's ability to detect early-stage lung cancer in animal models, monitored tumor mutational burden in patients with colorectal cancer and stratified patients with glioblastoma. SCOPE can expedite readouts, augmenting the clinical use of EVs in precision oncology.© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.