Tumor-derived extracellular vesicles (EVs) carry tumor-specific proteins and RNAs, thus becoming prevalent targets for early cancer diagnosis. However, low expression of EV cargos and insufficient diagnostic power of individual biomarkers hindered EVs application in clinical practice. Herein, we propose a multiplex Codetection platform of proteins and RNAs (Co-PAR) for EVs. Co-PAR adopted a pair of antibody-DNA probes to recognize the same target protein, which in turn formed a double-stranded DNA. Thus, the target protein could be quantified by detecting the double-stranded DNA via qPCR. Meanwhile, qRT-PCR simultaneously quantified the target RNAs. Thus, with a regular qPCR instrument, Co-PAR enabled the codetection of multiplex proteins and RNAs, with the sensitivity of 10² EVs/μL (targeting CD63) and 1 EV/μL (targeting snRNA U6). We analyzed the coexpressions of three protein markers (CD63, GPC-1, HER2) and three RNA markers (snRNA U6, GPC-1 mRNA, miR-10b) on EVs from three pancreatic cell lines and 30 human plasma samples using Co-PAR. The diagnostic accuracy of the 6-biomarker combination reached 92.9%, which was at least 6.2% higher than that of 3-biomarker combinations and at least 13.5% higher than that of 6 single biomarkers. Co-PAR, as a multiparameter detection platform for EVs, has great potential in early disease diagnosis.

中文翻译：

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细胞外囊泡上蛋白质和 RNA 的联合检测用于胰腺癌早期诊断

肿瘤来源的细胞外囊泡（EV）携带肿瘤特异性蛋白和RNA，因此成为早期癌症诊断的普遍靶标。然而，EV货物的低表达和单个生物标志物的诊断能力不足阻碍了EV在临床实践中的应用。在此，我们提出了用于 EV 的蛋白质和 RNA 多重编码检测平台 (Co-PAR)。 Co-PAR采用一对抗体-DNA探针来识别相同的目标蛋白，进而形成双链DNA。因此，可以通过 qPCR 检测双链 DNA 来定量靶蛋白。同时，qRT-PCR 同时对目标 RNA 进行定量。因此，使用常规 qPCR 仪器，Co-PAR 能够同时检测多重蛋白质和 RNA，灵敏度为 10 ² EVs/μL（针对 CD63）和 1 EV/μL（针对 snRNA U6）。我们使用 Co-PAR 分析了来自三种胰腺细胞系和 30 个人血浆样本的 EV 上三种蛋白质标记物（CD63、GPC-1、HER2）和三种 RNA 标记物（snRNA U6、GPC-1 mRNA、miR-10b）的共表达。 6个生物标志物组合的诊断准确率达到92.9%，比3个生物标志物组合至少高出6.2%，比6个单一生物标志物组合高至少13.5%。 Co-PAR作为EV多参数检测平台，在疾病早期诊断方面具有巨大潜力。