Bottom-Up Signal Boosting with Fractal Nanostructuring and Primer Exchange Reaction for Ultrasensitive Detection of Cancerous Exosomes.
Bottom-Up Signal Boosting with Fractal Nanostructuring and Primer Exchange Reaction for Ultrasensitive Detection of Cancerous Exosomes.
复制标题
DOI：
10.1021/acssensors.2c02819
复制
发表时间：
2023-02
期刊：
影响因子：
8.9
通讯作者：
M. Huang;Y-J Xiang;YU Chen;Hao Lu;Hui Zhang;Fengfei Liu;Xiaoling Qin;Xiaojie Qin;
中科院分区：
文献类型：
--
作者：
M. Huang;Y-J Xiang;YU Chen;Hao Lu;Hui Zhang;Fengfei Liu;Xiaoling Qin;Xiaojie Qin;
外切体是一种很有前景的癌症诊断生物标志物，但对肿瘤来源的外切体进行灵敏和准确的定量仍然是一个挑战。在这里，我们报道了一种超灵敏和特异的外切体传感器(NPExo)，它最初利用分层纳米结构阵列和引物交换反应(PER)来定量检测癌症外切体。NPExo使用一步电沉积法制备的高曲率纳米结构阵列(底部)来增强对目标外体的捕获。因此，免疫捕获的外切体提供了丰富的膜位置来插入大量密度比免疫配对高得多的胆固醇-DNA探针，这进一步允许基于PER的DNA延伸组装酶串联体(UP)以进行信号放大。这种自下而上的信号增强设计使NPExo具有高达75个粒子/毫升的超高灵敏度(即每10μL 1个外显体)和跨越6个数量级的宽动态范围。此外，我们的传感器还可以监测微小的胞外体表型转变，并显示出通过血液样本区分肝癌患者和健康捐赠者的高精度，表明NPExo在临床诊断中具有巨大的潜力。
Exosomes are emerging as promising biomarkers for cancer diagnosis, yet sensitive and accurate quantification of tumor-derived exosomes remains a challenge. Here, we report an ultrasensitive and specific exosome sensor (NPExo) that initially leverages hierarchical nanostructuring array and primer exchange reaction (PER) for quantitation of cancerous exosomes. This NPExo uses a high-curvature nanostructuring array (bottom) fabricated by single-step electrodeposition to enhance capturing of the target exosomes. The immuno-captured exosome thus provides abundant membrane sites to insert numerous cholesterol-DNA probes with a density much higher than that by immune pairing, which further allows PER-based DNA extension to assemble enzyme concatemers (up) for signal amplification. Such a bottom-up signal-boosting design imparts NPExo with ultrahigh sensitivity up to 75 particles/mL (i.e., <1 exosome per 10 μL) and a broad dynamic range spanning 6 orders of magnitude. Furthermore, our sensor allows monitoring subtle exosomal phenotypic transition and shows high accuracy in discrimination of liver cancer patients from healthy donors via blood samples, suggesting the great potential of NPExo as a promising tool in clinical diagnostics.