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Metal-ion-induced DNAzyme on magnetic beads for detection of lead(II) by using rolling circle amplification, glucose oxidase, and readout of pH changes

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Abstract

This work reports on a method for determination of lead(II) ion in environmental water samples. A Pb2+-specific DNAzyme immobilized on magnetic beads was coupled to rolling circle amplification (RCA) and pH meter-based readout. On addition of Pb2+ ion, it induces partial cleavage of the DNAzyme on the magnetic beads. The single-stranded DNA remaining on the magnetic beads is used as the primer to trigger the RCA reaction with the assistance of a circular DNA template, polymerase and dNTPs. This results in the formation of numerous oligonucleotide repeats on the magnetic bead. Subsequently, these repeats hybridize with glucose oxidase-labeled single-stranded DNA (GOx-ssDNA) to form a long concatamer containing tens to hundreds of GOx-ssDNA tandem repeats. The concatenated GOx molecules oxidize glucose, and this is accompanied by a drop in the local pH value. The pH values (vs. background signal) drop linearly when Pb2+ concentrations increase from 1.0–100 nM, and the detection limit is 0.91 nM. The method displays good reproducibility, high specificity and acceptable accuracy. It was applied to the analysis of spiked water samples, and results compared favorably with those obtained by ICP-MS.

Schematic presentation of metal-ion-induced DNAzyme on magnetic bead (MB) with rolling circle amplification (RCA) for pH meter-based detection of lead(II) ion and by using a glucose oxidase (GOx)-labeled probe.

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Scheme 1
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Acknowledgements

Authors acknowledged financial support from the National Natural Science Foundation of China (Grant nos.: 21675029 & 201874022) and the Health-Education Joint Research Project of Fujian Province (Grant no.: WKJ2016-2-15).

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Author notes

  1. Dianping Tang and Biyun Xia contributed equally to this work.

Authors and Affiliations

  1. Chongqing Key Laboratory of Environmental Materials & Remediation Technologies, Chongqing University of Arts and Sciences, Chongqing, 402160, People’s Republic of China

    Dianping Tang, Biyun Xia, Ying Tang & Jin Zhang

  2. Institute of Environmental and Analytical Science, School of Chemistry and Chemical Engineering, Henan University, Kaifeng, 475004, People’s Republic of China

    Qian Zhou

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  1. Dianping Tang
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  2. Biyun Xia
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  3. Ying Tang
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  4. Jin Zhang
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  5. Qian Zhou
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Correspondence to Dianping Tang or Qian Zhou.

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Tang, D., Xia, B., Tang, Y. et al. Metal-ion-induced DNAzyme on magnetic beads for detection of lead(II) by using rolling circle amplification, glucose oxidase, and readout of pH changes. Microchim Acta 186, 318 (2019). https://doi.org/10.1007/s00604-019-3454-1

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