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Ultrasensitive voltammetric determination of kanamycin using a target-triggered cascade enzymatic recycling couple along with DNAzyme amplification

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Abstract

The authors describe a label-free electrochemical aptasensor for ultrasensitive and highly specific detection of the antibiotic based on cascade enzymatic recycling couple with DNAzyme amplification. The assay involves two sequential reactions: the first reaction is a λ exonuclease-assisted cyclic digestion reaction triggered by target-aptamer binding. The second reaction is a nicking endonuclease-aided cyclic nicking reaction, which produces a large amount of G-rich nucleic acid segments. These form a G-quadruplex/hemin complex in the presence of K(I) ions and hemin. Because the G-quadruplex/hemin complex acts as a horseradish peroxidase-mimicking DNAzyme with excellent redox activity, the electrochemical signal transduction is accomplished due to the electroreduction of H2O2. It appears that this work is the first example that cascade enzymatic recycling coupled to DNAzyme amplification is used for antibiotic detection. The aptasensor was applied to the quantitation of kanamycin and gave a response that is linear in the 1 pM to 10 nM kanamycin concentration range, with a detection limit as low as 0.5 pM. The working voltage (vs. Ag/AgCl) at which data can be acquired best is −0.35 V. The assay offers the advantages of remarkably improved sensitivity, use of affordable instrumentation, and simplified operation without the need for electrochemical labeling or addition of labile reagents. Thus, cascade enzymatic recycling coupled to DNAzyme amplification represents a versatile platform for highly sensitive and specific antibiotics detection.

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Acknowledgements

This work was supported by NSFC (21405060, 1471644), Shandong Province Natural Science Funds for Distinguished Young Scholars (JQ201410), and Shandong Province Natural Science Funds (ZR2015CM027).

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Authors and Affiliations

  1. School of Biological Sciences and Technology, University of Jinan, Jinan, 250022, People’s Republic of China

    Cong Han, Hui Li, Chenggong Xu, Jingfeng Wang, Yu Wang & Jiadong Huang

  2. Jinan Maternity and Child Care Hospital, Jinan, 250022, People’s Republic of China

    Rongguo Li

  3. School of Resources and Environment, University of Jinan, Jinan, 250022, People’s Republic of China

    Su Liu

  4. Key Laboratory of Chemical Sensing & Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan, 250022, People’s Republic of China

    Jiadong Huang

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  1. Cong Han
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  2. Rongguo Li
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  3. Hui Li
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  4. Su Liu
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  7. Yu Wang
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Correspondence to Yu Wang.

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Han, C., Li, R., Li, H. et al. Ultrasensitive voltammetric determination of kanamycin using a target-triggered cascade enzymatic recycling couple along with DNAzyme amplification. Microchim Acta 184, 2941–2948 (2017). https://doi.org/10.1007/s00604-017-2311-3

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  • DOI: https://doi.org/10.1007/s00604-017-2311-3

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