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Fluorometric determination of nucleic acids based on the use of polydopamine nanotubes and target-induced strand displacement amplification

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Abstract

The authors describe a fluorometric method for the quantitation of nucleic acids by combining (a) cycled strand displacement amplification, (b) the unique features of the DNA probe SYBR Green, and (c) polydopamine nanotubes. SYBR Green undergoes strong fluorescence enhancement upon intercalation into double-stranded DNA (dsDNA). The polydopamine nanotubes selectively adsorb single-stranded DNA (ssDNA) and molecular beacons. In the absence of target DNA, the molecular beacon, primer and SYBR Green are adsorbed on the surface of polydopamine nanotubes. This results in quenching of the fluorescence of SYBR Green, typically measured at excitation/emission wavelengths of 488/518 nm. Upon addition of analyte (target DNA) and polymerase, the stem of the molecular beacon is opened so that it can bind to the primer. This triggers target strand displacement polymerization, during which dsDNA is synthesized. The hybridized target is then displaced due to the strand displacement activity of the polymerase. The displaced target hybridizes with another molecular beacon. This triggers the next round of polymerization. Consequently, a large amount of dsDNA is formed which is detected by addition of SYBR Green. Thus, sensitive and selective fluorometric detection is realized. The fluorescent sensing strategy shows very good analytical performances towards DNA detection, such as a wide linear range from 0.05 to 25 nM with a low limit of detection of 20 pM.

Schematic of a fluorometric strategy for highly sensitive and selective determination of nucleic acids by combining strand displacement amplification and the unique features of SYBR Green I (SG) and polydopamine nanotubes.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (21505122, 21205108), the Startup Research Fund of Zhengzhou University (1511316004), and the Outstanding Young Talent Research Fund of Zhengzhou University (1521316003, 1421316038).

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Authors and Affiliations

  1. College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou, 450001, People’s Republic of China

    Jia Ge, Dong-Mei Bai, Xin -Geng, Ya-Lei Hu, Ke Xing, Lin Zhang & Zhao-Hui Li

  2. Institute of Chemical Biology and Nanomedicine, Hunan University, Changsha, 410082, People’s Republic of China

    Qi-Yong Cai & Zhao-Hui Li

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  1. Jia Ge
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  2. Dong-Mei Bai
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  7. Lin Zhang
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  8. Zhao-Hui Li
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Correspondence to Jia Ge or Zhao-Hui Li.

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Ge, J., Bai, DM., -Geng, X. et al. Fluorometric determination of nucleic acids based on the use of polydopamine nanotubes and target-induced strand displacement amplification. Microchim Acta 185, 105 (2018). https://doi.org/10.1007/s00604-017-2632-2

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  • DOI: https://doi.org/10.1007/s00604-017-2632-2

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