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Highly efficient fluorescence sensing of kanamycin using Endo IV-powered DNA walker and hybridization chain reaction amplification

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Abstract

An ultrasensitive fluorescence sensing strategy for kanamycin (KANA) determination using endonuclease IV (Endo IV)-powered DNA walker, and hybridization chain reaction (HCR) amplification was reported. The sensing system consists of Endo IV-powered 3D DNA walker using for the specific recognition of KANA and the formation of the initiators, two metastable hairpin probes as the substrates of HCR and a tetrahydrofuran abasic site (AP site)-embeded fluorescence-quenched probe for fluorescence signal output. On account of this skilled design of sensing system, the specific binding between KANA and its aptamer activates DNA walker, in which the swing arm can move autonomously along the 3D track via Endo IV-mediated hydrolysis of the anchorages, inducing the formation of initiators that initiates HCR and the following Endo IV-assisted cyclic cleavage of fluorescence reporter probes. The use of Endo IV offers the advantages of simplified and accessible design without the need of specific sequence in DNA substrates. Under the optimal experimental conditions, the fluorescence biosensor shows excellent sensitivity toward KANA detection with a detection limit as low as 1.01 pM (the excitation wavelength is 486 nm). The practical applicability of this strategy is demonstrated by detecting KANA in spiked milk samples with recovery in the range of 98 to 102%. Therefore, this reported strategy might create an accurate and robust fluorescence sensing platform for trace amounts of antibiotic residues determination and related safety analysis.

Highly efficient fluorescence sensing of kanamycin using Endo IV-powered DNA Walker and hybridization chain, reaction amplification, Xiaonan Qu, Jingfeng Wang, Rufeng Zhang, Yihan Zhao, Shasha Li, Yu Wang, Su Liu*, Jiadong Huang, and Jinghua Yu, an ultrasensitive fluorescence sensing strategy for kanamycin determination using endonuclease IV-powered DNA walker, and hybridization chain reaction amplification is reported.

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Funding

This work was supported by National Natural Science Foundation of China (31471644), the Primary Research & Development Plan of Shandong Province (2017GSF220009) and the Program for Taishan Scholer of Shandong Province (TS201712048).

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Authors and Affiliations

  1. School of Water Conservancy and Environment, University of Jinan, Jinan, 250022, People’s Republic of China

    Xiaonan Qu, Rufeng Zhang & Su Liu

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

    Jingfeng Wang, Yihan Zhao, Shasha Li, Yu Wang & Jiadong Huang

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

    Jiadong Huang & Jinghua Yu

Authors
  1. Xiaonan Qu
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  2. Jingfeng Wang
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  3. Rufeng Zhang
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  4. Yihan Zhao
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  5. Shasha Li
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  6. Yu Wang
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  7. Su Liu
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  8. Jiadong Huang
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  9. Jinghua Yu
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Correspondence to Su Liu.

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Qu, X., Wang, J., Zhang, R. et al. Highly efficient fluorescence sensing of kanamycin using Endo IV-powered DNA walker and hybridization chain reaction amplification. Microchim Acta 187, 193 (2020). https://doi.org/10.1007/s00604-020-4167-1

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