Department of Pathology, The Affiliated People's Hospital of Ningbo University, No. 251 Baizhang Road, Yinzhou District, Ningbo, 315040, Zhejiang Province, China.
Biotechnol Lett. 2024 Dec;46(6):1049-1056. doi: 10.1007/s10529-024-03526-8. Epub 2024 Sep 12.
Precise identification of small extracellular vesicles (sEVs) is crucial for improving disease diagnosis and treatments, such as bladder cancer. However, accurate isolation and simultaneously quantification of sEVs remain a huge challenge. We have introduced a new technique that combines immobilization with aptamer-assisted dual cycle amplification to isolate and analyze sEVs with high sensitivity. In this method, the CD9 protein antibody is attached to the plate's surface for the initial identification of sEVs, while an aptamer probe is used to detect the exosomal surface protein CD63. We have created an sEVs-surface method that combines target recognition initiated signal recycling and rolling circle amplification (RCA) for signal amplification. This approach allows for the "AND" logic analysis of dual biomarkers, enabling both sEVs quantification and tracing. The proposed approach has a broad detection range and a low limit of detection. Moreover, the established method showed good stability in detecting sEVs with a low coefficient of variation. Our method can effectively isolate certain sEVs and accurately identify them, making it suitable for many uses in biological science, biomedical engineering, and personalized medicine.
精确识别小细胞外囊泡（sEVs）对于改善疾病诊断和治疗（如膀胱癌）至关重要。然而，sEVs 的精确分离和同时定量仍然是一个巨大的挑战。我们引入了一种新的技术，该技术结合了固定化和适体辅助双重循环扩增，以高灵敏度分离和分析 sEVs。在该方法中，CD9 蛋白抗体被附着到平板表面以初步鉴定 sEVs，而适体探针用于检测外泌体表面蛋白 CD63。我们创建了一种 sEVs-表面方法，该方法将目标识别引发的信号循环和滚环扩增（RCA）结合起来用于信号放大。这种方法允许对双生物标志物进行“与”逻辑分析，从而实现 sEVs 的定量和追踪。该方法具有较宽的检测范围和较低的检测限。此外，所建立的方法在检测 sEVs 时具有较低的变异系数，表现出良好的稳定性。我们的方法可以有效地分离特定的 sEVs 并准确识别它们，使其适用于生物科学、生物医学工程和个性化医学中的许多用途。
