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Electrochemical DNA biosensor based on MNAzyme-mediated signal amplification

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Abstract

The authors describe an electrochemical sensing strategy for highly sensitive and specific detection of target (analyte) DNA based on an amplification scheme mediated by a multicomponent nucleic acid enzyme (MNAzyme). MNAzymes were formed by multicomponent complexes which produce amplified “output” signals in response to specific “input” signal. In the presence of target nucleic acid, multiple partial enzymes (partzymes) oligonucleotides are assembled to form active MNAzymes. These can cleave H0 substrate into two pieces, thereby releasing the activated MNAzyme to undergo an additional cycle of amplification. Here, the two pieces contain a biotin-tagged sequence and a byproduct. The biotin-tagged sequences are specifically captured by the detection probes immobilized on the gold electrode. By employing streptavidinylated alkaline phosphatase as an enzyme label, an electrochemical signal is obtained. The electrode, if operated at a working potential of 0.25 V (vs. Ag/AgCl) in solution of pH 7.5, covers the 100 pM to 0.25 μM DNA concentration range, with a 79 pM detection limit. In our perception, the strategy introduced here has a wider potential in that it may be applied to molecular diagnostics and pathogen detection.

An electrochemical strategy for sequence-specific DNA detection based on multicomponent nucleic acid enzyme (MNAzyme) -mediated signal amplification.

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Acknowledgments

This work was funded by the National Natural Science Foundation of China (81371904), the Natural Science Foundation Project of CQ (CSTC2013jjB10019 and cstc2014kjrc-qnrc0012) and Application Development Plan Project of Chongqing (cstc2014yykfB10003).

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Authors and Affiliations

  1. Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China

    Wei Diao, Min Tang, Xiaojuan Ding, Ye Zhang, Jianru Yang, Wenbin Cheng, Fei Mo, Bo Wen, Lulu Xu & Yurong Yan

Authors
  1. Wei Diao
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  2. Min Tang
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  3. Xiaojuan Ding
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  4. Ye Zhang
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  5. Jianru Yang
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  6. Wenbin Cheng
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  7. Fei Mo
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  8. Bo Wen
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  9. Lulu Xu
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  10. Yurong Yan
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Corresponding author

Correspondence to Yurong Yan.

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Wei Diao and Min Tang contributed equally to this work.

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Diao, W., Tang, M., Ding, X. et al. Electrochemical DNA biosensor based on MNAzyme-mediated signal amplification. Microchim Acta 183, 2563–2569 (2016). https://doi.org/10.1007/s00604-016-1910-8

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  • DOI: https://doi.org/10.1007/s00604-016-1910-8

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