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A pH-responsive bioassay for sensitive colorimetric detection of adenosine triphosphate based on switchable DNA aptamer and metal ion–urease interactions

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Abstract

A facile and economic colorimetric strategy was designed for ATP detection by rationally using urease, a pH-responsive molecule, and a metal-mediated switchable DNA probe. By utilizing metal ions as a modulator of urease activity, the concentration of ATP is translated into pH change, which can be readily visualized by naked eye. An unmodified single-stranded DNA probe was designed, which consists of a target binding sequence and two flanked cytosine (C)-rich sequences. This C-rich single-stranded DNA can form a hairpin structure triggered by Ag+ ions via C-Ag+-C base mismatch. Upon introduction of ATP, Ag+-coordinated hairpin DNA structure will be broken and release the included Ag+, thus inhibiting the activity of urease. Conversely, urease can hydrolyze urea and raise pH value of the solution, resulting in the color change of the sensing solution. The proposed assay allows determination of ATP as low as 1.6 nM and shows a satisfactory result in human serum. Because of simple operation and low cost of this method, we believe it has a potential in point-of-care (POC) testing in resource-limited areas.

Graphical abstract

Schematic illustration of pH-responsive colorimetric sensor for ATP detection based on switchable DNA aptamer and metal ion–urease interactions

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Acknowledgments

The authors wish to express their gratitude to the staff at the Department of Oncology at the First Affiliated Hospital of Nan**g Medical University, Nan**g, China, for providing the human serum samples used for validation purposes. The authors gratefully acknowledge reagent support from Prof. Genxi Li.

Funding

This work was supported by the National Natural Science Foundation of China (Grant Nos. 81972484), The High-Level Innovation Team of Nan**g Medical University (JX102GSP201727), and The National Key Research and Development Program of China (ZDZX2017ZL-01).

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Authors and Affiliations

  1. Department of Oncology, The First Affiliated Hospital of Nan**g Medical University, Nan**g, 210029, People’s Republic of China

    Sha Zhu, Yiqi Yang & Yongmei Yin

  2. Department of Oncology, The Affiliated Wuxi No.2 People’s Hospital of Nan**g Medical University, Wuxi, 214000, People’s Republic of China

    Sha Zhu

  3. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nan**g University, Nan**g, 210093, People’s Republic of China

    Menglu Li & Yi Yang

  4. School of Food and Biological Engineering, Hefei University of Technology, Hefei, 230009, People’s Republic of China

    Chao Li

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  1. Sha Zhu
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  2. Yiqi Yang
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  3. Menglu Li
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  4. Yi Yang
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  5. Chao Li
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Correspondence to Chao Li or Yongmei Yin.

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The authors declare that they have no conflicts of interest.

Statement of ethical approval

All patient samples were acquired from Department of Oncology at the First Affiliated Hospital of Nan**g Medical University. The use of the discarded human serum samples was approved by the First Affiliated Hospital of Nan**g Medical University Institutional Review Board under protocol #2019-SRFA-132.

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Zhu, S., Yang, Y., Li, M. et al. A pH-responsive bioassay for sensitive colorimetric detection of adenosine triphosphate based on switchable DNA aptamer and metal ion–urease interactions. Anal Bioanal Chem 413, 1533–1540 (2021). https://doi.org/10.1007/s00216-020-03136-z

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  • DOI: https://doi.org/10.1007/s00216-020-03136-z

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