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Fluorometric aptamer based determination of adenosine triphosphate based on deoxyribonuclease I-aided target recycling and signal amplification using graphene oxide as a quencher

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Abstract

The authors describe an aptamer based fluorometric assay for the determination of ATP. It is based on deoxyribonuclease I-aided target recycling and signal amplification. The DNA probe consists of two regions (sequences) that represent the capture probe and the signal probe, respectively. In the absence of ATP, the probe is adsorbed by the surface of graphene oxide (GO) via π-stacking interactions. This results in quenching of the fluorescent label (carboxyfluorescein) and protects it from being cleaved by DNase. Upon adding ATP, the probe will be repelled by GO because ATP binds to the aptamer. This triggers an increase in fluorescence as measured at excitation/emission wavelengths of 480/514 nm. The detection limit is as low as 0.2 nM, and the calibration plot is linear in the 10 to 400 nM ATP concentration range. The assay is specific and sensitive, and in our perception has a large potential in terms of detecting other species including pathogenic microorganisms, small molecules, metal ions, and proteins.

Schematic of the fluorescent strategy for adenosine triphosphate assay by using aptamer-based target recognition and deoxyribonuclease I-aided signal amplification.

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Acknowledgements

We would like to thank National Natural Science Foundation of hunan province (2016JJ3098), Outstanding Youth Scientific Research Project Funded by Education department of Hunan Province (15B169), Science and technology Innovation Team in Colleges and Universities in Hunan Province《Chinese traditional medicine for treatment of infectious diseases》(No: 15, Grxjb-7), Doctor start-up Foundation of Hunan University of Chinese Medicine (9982-1001019), Key Subjects of Hunan University of Chinese Medicine《pathogenic biology》(NO.1) and Project Funded by Hunan Provincial Level Course《Immunology and pathogenic biology》(No. 48) for the financial support.

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  1. Yi Ning and Ke Wei contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology, The Medicine School of Hunan University of Chinese Medicine, Changsha, Hunan, 410208, People’s Republic of China

    Yi Ning, Ke Wei, Lijuan Cheng & Jue Hu

  2. Experimental Center of Pathogenic Microorganism and Immunology, The Chinese Medicine School of Hunan University of Chinese Medicine, Xueshi road 300, Changsha, Hunan, 410208, People’s Republic of China

    Qin Xiang

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  1. Yi Ning
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  2. Ke Wei
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Correspondence to Qin Xiang.

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Ning, Y., Wei, K., Cheng, L. et al. Fluorometric aptamer based determination of adenosine triphosphate based on deoxyribonuclease I-aided target recycling and signal amplification using graphene oxide as a quencher. Microchim Acta 184, 1847–1854 (2017). https://doi.org/10.1007/s00604-017-2194-3

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  • DOI: https://doi.org/10.1007/s00604-017-2194-3

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