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Highly sensitive DNA-based fluorometric mercury(II) bioassay based on graphene oxide and exonuclease III-assisted signal amplification

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Abstract

The article describes a fluorometric and sensitive assay for mercury(II) ions (Hg2+). It is based on the following scheme and experimental steps: (1) Hg2+ triggers the self-hybridization of thymine-rich ss-DNA labeled with a fluorescence tag to form a ds-DNA; (2) in the absence of Hg2+, labeled ss-DNA will be adsorbed on the surface of graphene oxide (GO) and its fluorescence is quenched; (3) the ds-DNA formed in the presence of Hg2+ is cleaved by the catalytic action of exonuclease III; (4) the cleaved labeled DNA fragments do not adsorb on the surface of GO, this resulting in an increase in fluorescence intensity. The induction of the process by Hg2+ leads to a strong amplification of fluorescence, while the fluorescence of uncleaved labeled ss-DNA is quenched because it is adsorbed on the surface of GO in the absence of Hg2+. This assay displays a detection limit of 0.1 nM (which is below the 10 nM upper limit in drinking water according to the US EPA and can be performed with 8 min.

The detection scheme is based on the finding that the fluorescence of ds-DNA formed in the presence of Hg(II) ions on the surface of graphene oxide is quenched. If the DNA is cleaved by exonuclease III, fragments will be desorbed and fluorescence pops up

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References

  1. Wang G, Xu G, Zhu Y, Zhang X (2014) A “turn-on” carbon nanotube-Ag nanoclusters fluorescent sensor for sensitive and selective detection of Hg2+ with cyclic amplification of exonuclease III activity. Chem Commun 50:747–750

    Article  CAS  Google Scholar 

  2. Tan D, He Y, Xing X, Zhao Y, Tang H, Pang D (2013) Aptamer functionalized gold nanoparticles based fluorescent probe for the detection of mercury (II) ion in aqueous solution. Talanta 113:26–30

    Article  CAS  Google Scholar 

  3. Wang R-Z, Zhou D-L, Huang H, Zhang M, Feng J-J, Wang A-J (2013) Water-soluble homo-oligonucleotide stabilized fluorescent silver nanoclusters as fluorescent probes for mercury ion. Microchim Acta 180:1287–1293

    Article  CAS  Google Scholar 

  4. Zhang L, Li T, Li B, Li J, Wang E (2010) Carbon nanotube-DNA hybrid fluorescent sensor for sensitive and selective detection of mercury(ii) ion. Chem Commun 46:1476–1478

    Article  CAS  Google Scholar 

  5. Huang J, Gao X, Jia J, Kim J-K, Li Z (2014) Graphene oxide-based amplified fluorescent biosensor for Hg2+ Detection through hybridization chain reactions. Anal Chem 86:3209–3215

    Article  CAS  Google Scholar 

  6. Li M, Zhou X, Ding W, Guo S, Wu N (2013) Fluorescent aptamer-functionalized graphene oxide biosensor for label-free detection of mercury(II). Biosens Bioelectron 41:889–893

    Article  Google Scholar 

  7. Li Y, Yan X-P, Dong L-M, Wang S-W, Jiang Y, Jiang D-Q (2005) Development of an ambient temperature post-column oxidation system for high-performance liquid chromatography on-line coupled with cold vapor atomic fluorescence spectrometry for mercury speciation in seafood. J Anal At Spectrom 20:467–472

    Article  Google Scholar 

  8. Pourreza N, Ghanemi K (2009) Determination of mercury in water and fish samples by cold vapor atomic absorption spectrometry after solid phase extraction on agar modified with 2-mercaptobenzimidazole. J Hazard Mater 161:982–987

    Article  CAS  Google Scholar 

  9. Zhu X, Alexandratos SD (2007) Determination of trace levels of mercury in aqueous solutions by inductively coupled plasma atomic emission spectrometry: elimination of the ‘memory effect’. Microchem J 86:37–41

    Article  CAS  Google Scholar 

  10. Hsu K-C, Lee C-F, Tseng W-C, Chao Y-Y, Huang Y-L (2014) Selective and eco-friendly method for determination of mercury(II) ions in aqueous samples using an on-line AuNPs-PDMS composite microfluidic device/ICP-MS system. Talanta 128:408–413

    Article  CAS  Google Scholar 

  11. Ono A, Togashi H (2004) Highly selective oligonucleotide-based sensor for Mercury(II) in aqueous solutions. Angew Chem Int Ed 43:4300–4302

    Article  CAS  Google Scholar 

  12. Miyake Y, Togashi H, Tashiro M, Yamaguchi H, Oda S, Kudo M, Tanaka Y, Kondo Y, Sawa R, Fujimoto T, Machinami T, Ono A (2006) MercuryII-mediated formation of Thymine − HgII − Thymine base pairs in DNA Duplexes. J Am Chem Soc 128:2172–2173

    Article  CAS  Google Scholar 

  13. Chung CH, Kim JH, Jung J, Chung BH (2013) Nuclease-resistant DNA aptamer on gold nanoparticles for the simultaneous detection of Pb2+ and Hg2+ in human serum. Biosens Bioelectron 41:827–832

    Article  CAS  Google Scholar 

  14. Du J, Liu M, Lou X, Zhao T, Wang Z, Xue Y, Zhao J, Xu Y (2012) Highly sensitive and selective chip-based fluorescent sensor for mercuric ion: development and comparison of turn-on and turn-off systems. Anal Chem 84:8060–8066

    Article  CAS  Google Scholar 

  15. He Y, Zhang X, Zeng K, Zhang S, Baloda M, Gurung AS, Liu G (2011) Visual detection of Hg2+ in aqueous solution using gold nanoparticles and thymine-rich hairpin DNA probes. Biosens Bioelectron 26:4464–4470

    Article  CAS  Google Scholar 

  16. Hu P, Jin L, Zhu C, Dong S (2011) A simple and sensitive fluorescent sensing platform for Hg2+ ions assay based on G-quenching. Talanta 85:713–717

    Article  CAS  Google Scholar 

  17. Guo S, Du D, Tang L, Ning Y, Yao Q, Zhang G-J (2013) PNA-assembled graphene oxide for sensitive and selective detection of DNA. Analyst 138:3216–3220

    Article  CAS  Google Scholar 

  18. Zhang Q, Kong D-M (2013) A general fluorescent sensor design strategy for “turn-on” activity detection of exonucleases and restriction endonucleases based on graphene oxide. Analyst 138:6437–6444

    Article  CAS  Google Scholar 

  19. Qi L, Zhao Y, Yuan H, Bai K, Zhao Y, Chen F, Dong Y, Wu Y (2012) Amplified fluorescence detection of mercury(ii) ions (Hg2+) using target-induced DNAzyme cascade with catalytic and molecular beacons. Analyst 137:2799–2805

    Article  CAS  Google Scholar 

  20. Xuan F, Luo X, Hsing IM (2013) Conformation-dependent Exonuclease III activity mediated by metal ions reshuffling on thymine-rich DNA duplexes for an ultrasensitive electrochemical method for Hg2+ detection. Anal Chem 85:4586–4593

    Article  CAS  Google Scholar 

  21. Wolfbeis OS (2013) Editorial: probes, sensors, and labels: why is real progress slow? Angew Chem Int Ed 52:9864–9865

    Article  CAS  Google Scholar 

  22. Xu Q, Cao A, Zhang L-F, Zhang C-Y (2012) Rapid and label-free monitoring of Exonuclease III-assisted target recycling amplification. Anal Chem 84:10845–10851

    Article  CAS  Google Scholar 

  23. Liu S, Wang C, Zhang C, Wang Y, Tang B (2013) Label-free and ultrasensitive electrochemical detection of nucleic acids based on autocatalytic and Exonuclease III-assisted target recycling strategy. Anal Chem 85:2282–2288

    Article  CAS  Google Scholar 

  24. Chen H, Wang J, Liang G, Zhang P, Kong J (2012) A novel exonuclease III aided amplification method for sensitive nucleic acid detection based on single walled carbon nanotube induced quenching. Chem Commun 48:269–271

    Article  CAS  Google Scholar 

  25. Chen C, Xiang X, Liu Y, Zhou G, Ji X, He Z (2014) Dual-color determination of protein via terminal protection of small-molecule-linked DNA and the enzymolysis of exonuclease III. Biosens Bioelectron 58:205–208

    Article  CAS  Google Scholar 

  26. Cheng G, Wang Z-G, Liu Y-L, Zhang J-L, Sun D-H, Ni J-Z (2012) A graphene-based multifunctional affinity probe for selective capture and sequential identification of different biomarkers from biosamples. Chem Commun 48:10240–10242

    Article  CAS  Google Scholar 

  27. Xuan F, Luo X, Hsing IM (2012) Ultrasensitive solution-phase electrochemical molecular beacon-based DNA detection with signal amplification by Exonuclease III-assisted target recycling. Anal Chem 84:5216–5220

    Article  CAS  Google Scholar 

  28. Cui X, Zhu L, Wu J, Hou Y, Wang P, Wang Z, Yang M (2015) A fluorescent biosensor based on carbon dots-labeled oligodeoxyribonucleotide and graphene oxide for mercury (II) detection. Biosens Bioelectron 63:506–512

    Article  CAS  Google Scholar 

  29. Jiang Y, Tian J, Hu K, Zhao Y, Zhao S (2014) Sensitive aptamer-based fluorescence polarization assay for mercury(II) ions and cysteine using silver nanoparticles as a signal amplifier. Microchim Acta 181:1423–1430

    Article  CAS  Google Scholar 

  30. Deng L, Zhou Z, Li J, Li T, Dong S (2011) Fluorescent silver nanoclusters in hybridized DNA duplexes for the turn-on detection of Hg2+ ions. Chem Commun 47:11065–11067

    Article  CAS  Google Scholar 

  31. Guo L, Yin N, Nie D et al (2011) An ultrasensitive electrochemical sensor for the mercuric ion via controlled assembly of SWCNTs. Chem Commun 47:10665–10667

    Article  CAS  Google Scholar 

  32. Hao Y, Guo Q, Wu H et al (2014) Amplified colorimetric detection of mercuric ions through autonomous assembly of G-quadruplex DNAzyme nanowires. Biosens Bioelectron 52:261–264

    Article  CAS  Google Scholar 

  33. Wang G, Huang H, Zhang X et al (2012) Electrically contacted enzyme based on dual hairpin DNA structure and its application for amplified detection of Hg2+. Biosens Bioelectron 35:108–114

    Article  CAS  Google Scholar 

  34. Xue X, Wang F, Liu X (2008) One-step, room temperature, colorimetric detection of Mercury (Hg2+) using DNA/nanoparticle conjugates. J Am Chem Soc 130:3244–3245

    Article  CAS  Google Scholar 

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Acknowledgments

The authors acknowledge the support of Natural Science Foundation of China (Nos. 21275040 and 21475034), and the Natural Science Foundation of Hubei Province (No. 2013CFA061).

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Authors and Affiliations

  1. School of Laboratory Medicine, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan, 430065, China

    Yulin Zhang, Lina Tang, Fan Yang, Zhongyue Sun & Guo-Jun Zhang

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  1. Yulin Zhang
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  2. Lina Tang
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  3. Fan Yang
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  4. Zhongyue Sun
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  5. Guo-Jun Zhang
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Correspondence to Guo-Jun Zhang.

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Zhang, Y., Tang, L., Yang, F. et al. Highly sensitive DNA-based fluorometric mercury(II) bioassay based on graphene oxide and exonuclease III-assisted signal amplification. Microchim Acta 182, 1535–1541 (2015). https://doi.org/10.1007/s00604-015-1482-z

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  • DOI: https://doi.org/10.1007/s00604-015-1482-z

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