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Aptamer-based fluorometric determination of ATP by using target-cycling strand displacement amplification and copper nanoclusters

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Abstract

A fluorescence aptasensor is described that combines target-cycling strand displacement amplification (TCSDA) and synthesis of copper nanoclusters (CuNCs) templated with double-stranded DNA (dsDNA). Specifically, the detection scheme was applied to the determination of adenosine-5′-triphosphate (ATP) via target-induced structure switching design. The binding of an aptamer hairpin probe (AP) to ATP induces a structural switch from a hairpin shape to an open conformation. This facilitates hybridization with a primer and triggers a TCSDA reaction. This amplification step produces a large quantity of dsDNA that can directly act as a template for the synthesis of fluorescent CuNCs, thereby producing a strong red fluorescence (with excitation/emission maxima at 340/598 nm) that can be used to quantify ATP. The assay has a dynamic range that extends over 4 decades (from 0.01 nM to 100 nM) and a 5 pM detection limit. Conceivably, this detection scheme is applicable to numerous other analytes for which suitable aptamers are available.

Schematic of an ATP assay (with a 5 pM detection limit) that is taking advantages of (a) target-cycling strand displacement amplification, and (b) of red fluorescent copper nanoclusters. KF polymerase: Klenow fragment of polymerase

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Scheme 1
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Acknowledgements

This work was financially supported by the National Key Basic Research Program (2011CB911000), National Natural Science Foundation of China (21527810, 21575036, 21190041, 21521063, 21505041) and Graduate Student Research Innovation Project of Hunan Province (CX2016B118).

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Authors and Affiliations

  1. State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, Hunan, 410082, People’s Republic of China

    Yu-Min Wang, Jin-Wen Liu, Lu-Ying Duan & Jian-Hui Jiang

  2. Guangxi Collaborative Innovation Center for Biomedicine and Guangxi Key Laboratory of Regenerative Medicine, School of Preclinical Medicine, Guangxi Medical University, Nanning, Guangxi, 530021, People’s Republic of China

    Si-Jia Liu

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  1. Yu-Min Wang
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  2. Jin-Wen Liu
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Correspondence to Si-Jia Liu or Jian-Hui Jiang.

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Wang, YM., Liu, JW., Duan, LY. et al. Aptamer-based fluorometric determination of ATP by using target-cycling strand displacement amplification and copper nanoclusters. Microchim Acta 184, 4183–4188 (2017). https://doi.org/10.1007/s00604-017-2337-6

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