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Rapid and efficient isolation and detection of extracellular vesicles from plasma for lung cancer diagnosis

机译:肺癌诊断等血浆中细胞外囊的快速有效的分离和检测

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Extracellular vesicles (EVs) are cell-derived nanoscale vesicles that provide promising biomarkers for the non-invasive diagnosis of cancer because they carry important cancer-related DNA, RNA and protein biomarkers. However, the clinical application of EVs is limited by tedious and non-standardized isolation methods that require bulky instrumentation. Here, we propose an easy-to-operate, simple dielectrophoretic (DEP) method for EV isolation with higher recovery efficiency (83%) and higher purity than ultracentrifugation (UC). The DEP chip reduces the isolation procedure from 8 h to 30 min. To facilitate subsequent analysis, our DEP chip achieved integration of EV isolation and in situ lysis of EVs for the first time. Our chip also achieved on-chip siRNA delivery to EVs isolated by DEP. We found that EVs isolated from the plasma of lung cancer patients contained higher levels of miR-21, miR-191 and miR-192 compared to those from healthy people. With on-chip detection, EGFR in EVs could distinguish lung cancer patients from healthy people. Overall, this study provides an efficient and practical approach to the isolation and detection of EVs, which could be used for the early diagnosis of lung cancer.
机译:细胞外囊泡(EVS)是细胞衍生的纳米级囊泡,其为癌症的非侵入性诊断提供了有前途的生物标志物,因为它们携带重要的癌症相关的DNA,RNA和蛋白质生物标志物。然而,EVS的临床应用受到需要庞大仪器的乏味和非标准化隔离方法的限制。在这里,我们提出了一种易于操作,简单的介电电泳(DEP)方法,用于EV隔离,具有较高的恢复效率(& 83%)和比超速离心(UC)更高的纯度。 DEP芯片将8小时降低到30分钟。为了促进随后的分析,我们的DEP芯片首次实现了EV分离和EVS原位裂解的整合。我们的芯片还实现了片上siRNA交付给Deps隔离的EV。我们发现,与来自健康人的人相比,从肺癌患者的血浆中分离的EVS包含更高水平的MiR-21,MiR-191和MiR-192。随着片上检测,EVS中的EGFR可以区分来自健康人的肺癌患者。总体而言,本研究提供了对EV的分离和检测的有效和实用的方法,可用于肺癌的早期诊断。

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