Li Chao, Guo Long, Sang Xiaoqiang, Jiang Xiaoli, Wang Hanwen, Qin Panzhu, Huang Lin
School of Life Sciences, Anhui Medical University, Hefei, Anhui, 230032, China.
Department of Nutrition and Food Hygiene, School of Public Health, Anhui Medical University, Hefei, 230032, Anhui, China.
Talanta. 2023 Jun 1;258:124453. doi: 10.1016/j.talanta.2023.124453. Epub 2023 Mar 13.
Exosomes are one of the most promising biomarkers for tumor diagnosis and prognosis. Therefore, the development of convenient and sensitive exosome sensing strategies is of great significance. Herein, we integrated aptamer-based spherical nucleic acids (SNAs) and hybridization chain reaction (HCR) into a colorimetric aptasensor platform and applied it to the detection of exosomes. In this design, the CD63-specific aptamer pre-immobilized on the microplate was used to capture target exosomes, while the SNAs conjugated with nucleolin-specific aptamer and trigger probe H1 were designed for amplifying signal. In the presence of target exosomes, the SNAs can be attached to the microplate by the bridge effect of exosomes, resulting in the trigger of HCR. This process is accompanied by the formation of abundant G-quadruplex/hemin DNAzyme, enabling the visual quantitative analysis of exosomes. Featured with the dual amplification of SNAs and HCR, the proposed aptasensor achieved a considerable detection limit of 50 particles/μL. The practicability of this method was further verified by testing the different clinical samples. Given the ability of the aptasensor to visually detect exosomes in scenarios lacking instruments and resources, we believe that the aptasensor can be serve as a potential on-site test for liquid biopsy.
外泌体是肿瘤诊断和预后最有前景的生物标志物之一。因此,开发便捷、灵敏的外泌体传感策略具有重要意义。在此,我们将基于适配体的球形核酸(SNA)和杂交链式反应(HCR)整合到比色适配体传感器平台中,并将其应用于外泌体的检测。在该设计中,预先固定在微孔板上的CD63特异性适配体用于捕获目标外泌体,而与核仁素特异性适配体和触发探针H1偶联的SNA则用于信号放大。在存在目标外泌体的情况下,SNA可通过外泌体的桥连作用附着于微孔板上,从而触发HCR。这一过程伴随着大量G-四链体/血红素DNAzyme的形成,实现了外泌体的可视化定量分析。该适配体传感器具有SNA和HCR的双重放大作用,实现了50个颗粒/μL的可观检测限。通过检测不同临床样本进一步验证了该方法实用性。鉴于该适配体传感器能够在缺乏仪器和资源的情况下对外泌体进行可视化检测,我们认为该适配体传感器可作为液体活检的潜在现场检测方法。