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A fluorescent immunosensor for determination and imaging of circulating tumor cells based on a bifunctional DNA nanomachine

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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 toehold-mediated bifunctional DNA nanomachine was demonstrated by the specific gene mutation detection in single-cell analysis.

Schematic illustration of the fluorescent immunosensor for determination and imaging of circulating tumor cells. The method is based on aptamer-based recognition and toehold-mediated bifunctional DNA nanomachine.

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Funding

This study was supported by the National Natural Science Foundation of China (81802114 and 81871735) and the Science and Technology Planning Project of Guangdong Province (2017A020215123).

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Author notes

  1. Ye Zhang and Shihua Luo contributed equally to this work.

Authors and Affiliations

  1. Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, 510515, Guangzhou, Guangdong Province, People’s Republic of China

    Ye Zhang, Shihua Luo, Bo Situ, Xinyi Ye, Yifang Huang, Bo Li, Xiujuan Jiang, Lei Zheng & Xiaohui Yan

  2. Guangdong Engineering and Technology Research Center for Rapid Diagnostic Biosensors, Nanfang Hospital, Southern Medical University, 510515, Guangzhou, People’s Republic of China

    Ye Zhang, Shihua Luo, Bo Situ, Xinyi Ye, Yifang Huang, Bo Li, Xiujuan Jiang & Lei Zheng

  3. The Center for Clinical Molecular Medical Detection, The First Affiliated Hospital of Chongqing Medical University, 400016, Chongqing, People’s Republic of China

    Xueping Chen

  4. Research Center of Clinical Medicine, Nanfang Hospital, Southern Medical University, 510515, Guangzhou, People’s Republic of China

    Xiaohui Yan

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  1. Ye Zhang
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Correspondence to Lei Zheng or Xiaohui Yan.

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Zhang, Y., Luo, S., Situ, B. et al. A fluorescent immunosensor for determination and imaging of circulating tumor cells based on a bifunctional DNA nanomachine. Microchim Acta 187, 259 (2020). https://doi.org/10.1007/s00604-020-4205-z

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