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Signal-amplification detection of small molecules by use of Mg2+- dependent DNAzyme

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Abstract

Because small molecules can be beneficial or toxic in biology and the environment, specific and sensitive detection of small molecules is one of the most important objectives of the scientific community. In this study, new signal amplification assays for detection of small molecules based on Mg2+-dependent DNAzyme were developed. A cleavable DNA substrate containing a ribonucleotide, the ends of which were labeled with black hole quencher (BHQ) and 6-carboxyfluorescein (FAM), was used for fluorescence detection. When the small molecule of interest is added to the assay solution, the Mg2+-dependent DNAzyme is activated, facilitating hybridization between the Mg2+-dependent DNAzyme and the DNA substrate. Binding of the substrate to the DNAzyme structure results in hydrolytic cleavage of the substrate in the presence of Mg2+ ions. The fluorescence signal was amplified by continuous cleavage of the enzyme substrate. Ochratoxin A (OTA) and adenosine triphosphate (ATP) were used as model analytes in these experiments. This method can detect OTA specifically with a detection limit as low as 140 pmol L−1 and detect ATP specifically with a detection limit as low as 13 nmol L−1. Moreover, this method is potentially extendable to detection of other small molecules which are able to dissociate the aptamer from the DNAzyme, leading to activation of the DNAzyme.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (nos 21190040 and 21275137), Start Funding from Changchun Institute of Applied Chemistry (CAS), and Supporting Funding for Creative Young Scientists (CAS).

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Authors and Affiliations

  1. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China

    Zhijun Guo, Jiahai Wang & Erkang Wang

  2. Department of Food Science and Engineering, Yanbian University, YanJi, 133000, China

    Zhijun Guo

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  1. Zhijun Guo
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Correspondence to Jiahai Wang or Erkang Wang.

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Guo, Z., Wang, J. & Wang, E. Signal-amplification detection of small molecules by use of Mg2+- dependent DNAzyme. Anal Bioanal Chem 405, 4051–4057 (2013). https://doi.org/10.1007/s00216-013-6788-2

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  • DOI: https://doi.org/10.1007/s00216-013-6788-2

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