Department of Chemistry and Shanghai Stomatological Hospital, Fudan University, Shanghai, 200000, China; Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei, 230032, China.
Department of Chemistry and Shanghai Stomatological Hospital, Fudan University, Shanghai, 200000, China.
Biosens Bioelectron. 2022 Dec 1;217:114709. doi: 10.1016/j.bios.2022.114709. Epub 2022 Sep 11.
Osteosarcoma is one of the most frequent primary sarcoma of bone among adolescents. Early diagnosis of osteosarcoma is the key factor to achieve high survival rate of patients. Nevertheless, traditional histological biopsy is highly invasive and associated with the risk of arousing tumor spread. Herein, we develop a method integrating microfluidics and surface-enhanced Raman spectroscopy (SERS) to isolate plasma-derived exosomes and profile multiple exosomal biomarkers for the diagnosis of osteosarcoma. The method showed highly efficient isolation of exosomes directly from human plasma and can profile exosomes based on protein biomarkers, with the detection limit down to 2 exosomes per μL. The whole assay can be performed in 5 h and only consumed 50 μL of plasma for one analysis. With the method, we analyzed the level of three protein biomarkers, i.e., CD63, vimentin (VIM) and epithelial cell adhesion molecule (EpCAM), on plasma-derived exosomes from 20 osteosarcoma patients and 20 heathy controls. Significantly higher levels of CD63, VIM and EpCAM were observed on plasma exosomes from the osteosarcoma patients compared to the healthy controls. Based on the level of the exosomal biomarkers, a classification model was built for the rapid diagnosis of osteosarcoma, with the sensitivity, specificity and accuracy of 100%, 90% and 95%, respectively. The proposed method does not require complex operations nor expensive equipment, and has great promise in clinical diagnosis of cancer as a liquid biopsy technique.
骨肉瘤是青少年中最常见的原发性骨肉瘤之一。骨肉瘤的早期诊断是提高患者生存率的关键因素。然而，传统的组织学活检具有很强的侵入性，并伴有引起肿瘤扩散的风险。在此，我们开发了一种结合微流控和表面增强拉曼光谱（SERS）的方法，用于分离血浆来源的外泌体并分析多种外泌体生物标志物，以诊断骨肉瘤。该方法能够高效地直接从人血浆中分离出外泌体，并能够基于蛋白质生物标志物对其进行分析，检测限低至 2 个外泌体/μL。整个检测过程可以在 5 小时内完成，每次分析仅需消耗 50 μL 的血浆。利用该方法，我们分析了 20 名骨肉瘤患者和 20 名健康对照者血浆来源外泌体中三种蛋白质生物标志物（即 CD63、波形蛋白（VIM）和上皮细胞黏附分子（EpCAM））的水平。与健康对照组相比，骨肉瘤患者血浆外泌体中 CD63、VIM 和 EpCAM 的水平明显升高。基于外泌体标志物的水平，我们建立了一个快速诊断骨肉瘤的分类模型，其灵敏度、特异性和准确性分别为 100%、90%和 95%。该方法不需要复杂的操作或昂贵的设备，有望成为癌症的液体活检技术，用于临床诊断。
