A fluorescent immunosensor for determination and imaging of circulating tumor cells based on a bifunctional DNA nanomach
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ORIGINAL PAPER
A fluorescent immunosensor for determination and imaging of circulating tumor cells based on a bifunctional DNA nanomachine Ye Zhang 1,2 & Shihua Luo 1,2 & Bo Situ 1,2 & Xinyi Ye 1,2 & Yifang Huang 1,2 & Bo Li 1,2 & Xiujuan Jiang 1,2 & Xueping Chen 3 & Lei Zheng 1,2 & Xiaohui Yan 1,4 Received: 3 February 2020 / Accepted: 28 February 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract A fluorescent platform was developed for the determination and visualization of circulating tumor cells by a toehold-mediated bifunctional DNA nanomachine. In the presence of target tumor cells, the DNA nanomachine was activated. Multiple DNA products were formed, including dendritic DNA products and double-strand DNA products. Dendritic DNA products bound to their target cells for the visualization, while double-strand DNA products were released for the determination of tumor cells. At fluorescence excitation and emission wavelengths of 530 and 550 nm, this method could detect as low as 43 cells/mL (S/N = 3) with a linear range of 100 to 10,000 cells/mL. In clinical hydrothorax samples, this platform exhibited high reliability with a recovery of 93 to 116%. At the fluorescence excitation and emission wavelengths of 490 and 515 nm, the specificity and biocompatibility of this method were further verified by tumor cells imaging. Furthermore, the robustness of the toeholdmediated bifunctional DNA nanomachine was demonstrated by the specific gene mutation detection in single-cell analysis. Keywords Fluorescent platform . Toehold-mediated . Visualization . Gene mutation detection
Introduction The analysis of circulating tumor cells (CTCs) plays essential roles in early clinical diagnosis, evaluation of cancer Ye Zhang and Shihua Luo contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-020-4205-z) contains supplementary material, which is available to authorized users. * Lei Zheng [email protected] * Xiaohui Yan [email protected] 1
Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, 510515 Guangzhou, Guangdong Province, People’s Republic of China
2
Guangdong Engineering and Technology Research Center for Rapid Diagnostic Biosensors, Nanfang Hospital, Southern Medical University, 510515 Guangzhou, People’s Republic of China
3
The Center for Clinical Molecular Medical Detection, The First Affiliated Hospital of Chongqing Medical University, 400016 Chongqing, People’s Republic of China
4
Research Center of Clinical Medicine, Nanfang Hospital, Southern Medical University, 510515 Guangzhou, People’s Republic of China
recurrence, and individualized anti-cancer treatment [1–5]. The scarcity of CTCs in clinical specimens makes their analysis challenging. To address this issue, various strategies, including flow cytometry [6–8], polymerase chain reaction [9–11], electrochemical methods [12–16], and fluorescent spectrometry [17–20], have been developed. Among these methods, fluorescent sp
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