Microchimica Acta

, 185:503 | Cite as

A voltammetric assay for microRNA-25 based on the use of amino-functionalized graphene quantum dots and ss- and ds-DNAs as gene probes

  • Azam Akbarnia
  • Hamid R. ZareEmail author
Original Paper


The authors describe a DNA based voltammetric assay for the cancer biomarker microRNA-25. A glassy carbon electrode (GCE) was modified with amino-functionalized graphene quantum dots and used as an amplifier of electrochemical signals. p-Biphenol is introduced as a new electroactive probe with a fairly low working potential of 0.3 V (vs. Ag/AgCl). The stages of fabricating the electrode were characterized by cyclic voltammetry and electrochemical impedance spectroscopy. ss-Probe DNA was immobilized on the modified GCE and then exposed to a sample containing microRNA-25. The results indicated that the electrode can distinguish complementary microRNA-25 from a single-base mismatch. The increase in the electrochemical response of PBP and the positive shift in the potential peak indicate that PBP is intercalated between two strands. Under optimized experimental conditions, the current of the electrode increases linearly with the logarithm of the microRNA-25 concentration in the range from 0.3 nM to 1.0 μM, and the detection limit is 95.0 pM. The assay was successfully employed to the determination of microRNA-25 in spiked human plasma.

Graphical abstract

A novel electrochemical nanogenosensor is introduced for simple and sensitive determination of microRNA-25, as a biomarker, based on amino-functionalized graphene quantum dots (as a surface modifier) and p-biphenol (as an electroactive label).


Pyrolysis Citric acid p-Biphenol Amino-functionalized graphene quantum dots microRNA-25 Electroactive label 


Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_3037_MOESM1_ESM.docx (556 kb)
ESM 1 (DOCX 556 kb)


  1. 1.
    Li L, Xiao B, Tong H, Xie F, Zhang Z, Xiao GG (2012) Regulation of breast cancer tumorigenesis and metastasis by miRNAs. Expert Rev Proteomics 9:615–625CrossRefGoogle Scholar
  2. 2.
    Chen Z-Y, Chen X, Wang Z-X (2016) The role of microRNA-196a in tumorigenesis, tumor progression, and prognosis. Tumor Biol 37:15457–15466CrossRefGoogle Scholar
  3. 3.
    Tüfekci KU, Öner MG, Meuwissen RLJ, Genç Ş (2014) The role of microRNAs in human diseases. miRNomics: MicroRNA biology and computational analysis. In: Methods in Molecular Biology (Methods and Protocols), vol 1107. Humana Press, TotowaGoogle Scholar
  4. 4.
    Feng K, Liu J, Deng L, Yu H, Yang M (2018) Amperometric detection of microRNA based on DNA-controlled current of a molybdophosphate redox probe and amplification via hybridization chain reaction. Microchim Acta 185:28CrossRefGoogle Scholar
  5. 5.
    Liu L, Jiang S, Wang L, Zhang Z, Xie G (2015) Direct detection of microRNA-126 at a femtomolar level using a glassy carbon electrode modified with chitosan, graphene sheets, and a poly (amidoamine) dendrimer composite with gold and silver nanoclusters. Microchim Acta 182:77–846CrossRefGoogle Scholar
  6. 6.
    Zhou Y, Yin H, Li J, Li B, Li X, Ai S, Zhang X (2016) Electrochemical biosensor for microRNA detection based on poly (U) polymerase mediated isothermal signal amplification. Biosens Bioelectron 79:79–85CrossRefGoogle Scholar
  7. 7.
    Wu T, Chen W, Kong D, Li X, Lu H, Liu S, Wang J, du L, Kong Q, Huang X, Lu Z (2015) miR-25 targets the modulator of apoptosis 1 gene in lung cancer. Carcinogenesis 36:925–935CrossRefGoogle Scholar
  8. 8.
    Zhang H, Zuo Z, Lu X, Wang L, Wang H, Zhu Z (2012) MiR-25 regulates apoptosis by targeting bim in human ovarian cancer. Oncol Rep 27:594–598PubMedGoogle Scholar
  9. 9.
    Esposito F, Tornincasa M, Pallante P, Federico A, Borbone E, Pierantoni GM, Fusco A (2012) Down-regulation of the mir-25 and mir-30d contributes to the development of anaplastic thyroid carcinoma targeting the polycomb protein EZH2. J Clin Endocrinol Metab 97(5):E710–E718CrossRefGoogle Scholar
  10. 10.
    Azimzadeh M, Rahaie M, Nasirizadeh N, Naderi-Manesh H (2015) Application of oracet blue in a novel and sensitive electrochemical biosensor for the detection of microRNA. Anal Methods 7:9495–9503CrossRefGoogle Scholar
  11. 11.
    Rafiee-Pour H-A, Behpour M, Keshavarz M (2016) A novel label-free electrochemical miRNA biosensor using methylene blue as redox indicator: application to breast cancer biomarker miRNA-21. Biosens Bioelectron 77:202–207CrossRefGoogle Scholar
  12. 12.
    Xia N, Wang X, Deng D, Wang G, Zhai H, Li S-J (2013) Label-free electrochemical sensor for MicroRNAs detection with ferroceneboronic acids as redox probes. Int J Electrochem Sci 8:9714–9722Google Scholar
  13. 13.
    Murakami Y, Ishii H, Hoshina S, Takada N, Ueki A, Tanaka S, Kadoma Y, Ito S, Machino M, Fujisawa S (2009) Antioxidant and cyclooxygenase-2-inhibiting activity of 4,4′-biphenol, 2,2′-biphenol and phenol. Anticancer Res 29:2403–2410PubMedGoogle Scholar
  14. 14.
    Deep A, Jain S, Sharma PC, Verma P, Kumar M, Dora CP (2010) Design and biological evaluation of biphenyl-4-carboxylic acid hydrazide-hydrazone for antimicrobial activity. Acta Pol Pharm 67:255–259PubMedGoogle Scholar
  15. 15.
    Belsare PU, Zade AB (2013) Synthesis, characterization and thermal study of 2, 2′-biphenol-tetraethylenepentamine-formaldehyde terpolymer resin. Der Pharma Chemica 5(4):325–334Google Scholar
  16. 16.
    Fraser DM (1994) Biphenol as an electron transfer mediator for glucose oxidase. Anal Lett 27:2039–2053CrossRefGoogle Scholar
  17. 17.
    Shayani-Jam H, Nematollahi D (2011) Electrochemically mediated oxidation of glutathione and N-acetylcysteine with 4, 4′-biphenol. Electrochim Acta 56:9311–9316CrossRefGoogle Scholar
  18. 18.
    Laurenti M, Paez-Perez M, Algarra M, Alonso-Cristobal P, Lopez-Cabarcos E, Mendez-Gonzalez D, Rubio-Retama J (2016) Enhancement of the upconversion emission by visible-to-near-infrared fluorescent graphene quantum dots for miRNA detection. ACS Appl Mater Interfaces 8:12644–12651CrossRefGoogle Scholar
  19. 19.
    Huang S, Qiu H, Zhu F, Lu S, Xiao Q (2015) Graphene quantum dots as on-off-on fluorescent probes for chromium (VI) and ascorbic acid. Microchim Acta 182:1723–1731CrossRefGoogle Scholar
  20. 20.
    Zhang T, Zhao H, Fan G, Li Y, Li L, Quan X (2016) Electrolytic exfoliation synthesis of boron doped graphene quantum dots: a new luminescent material for electrochemiluminescence detection of oncogene microRNA-20a. Electrochim Acta 190:1150–1158CrossRefGoogle Scholar
  21. 21.
    Dong Y, Shao J, Chen C, Li H, Wang R, Chi Y, Lin X, Chen G (2012) Blue luminescent graphene quantum dots and graphene oxide prepared by tuning the carbonization degree of citric acid. Carbon 50:4738–4743CrossRefGoogle Scholar
  22. 22.
    Sun H, Gao N, Wu L, Ren J, Wei W, Qu X (2013) Highly Photoluminescent amino-functionalized graphene quantum dots used for sensing copper ions. Chemistry Eur J 19:13362–13368CrossRefGoogle Scholar
  23. 23.
    Panagopoulou MA, Stergiou DV, Roussis IG, Prodromidis MI (2010) Impedimetric biosensor for the assessment of the clotting activity of rennet. Anal Chem 82:8629–8636CrossRefGoogle Scholar
  24. 24.
    Suni II (2008) Impedance methods for electrochemical sensors using nanomaterials. TrAC Trends Anal Chem 27:604–611CrossRefGoogle Scholar
  25. 25.
    Razmi H, Mohammad-Rezaei R (2013) Graphene quantum dots as a new substrate for immobilization and direct electrochemistry of glucose oxidase: application to sensitive glucose determination. Biosens Bioelectron 41:498–504CrossRefGoogle Scholar
  26. 26.
    Carter MT, Rodriguez M, Bard AJ (1989) Voltammetric studies of the interaction of metal chelates with DNA. 2. Tris-chelated complexes of cobalt (III) and iron (II) with 1, 10-phenanthroline and 2, 2′-bipyridine. J Am Chem Soc 111:8901–8911CrossRefGoogle Scholar
  27. 27.
    Miller JN, Miller JC (2000) Statistics and Chemometrics for analytical chemistry, 4th Pearson education limited, EssexGoogle Scholar
  28. 28.
    Li F, Peng J, Wang J, Tang H, Tan L, Xie Q, Yao S (2014) Carbon nanotube-based label-free electrochemical biosensor for sensitive detection of miRNA-24. Biosens Bioelectron 54:158–164CrossRefGoogle Scholar
  29. 29.
    Liu S, Su W, Li Z, Ding X (2015) Electrochemical detection of lung cancer specific microRNAs using 3D DNA origami nanostructures. Biosens Bioelectron 71:57–61CrossRefGoogle Scholar
  30. 30.
    Chen Y, Xiang Y, Yuan R, Chai Y (2015) Intercalation of quantum dots as the new signal acquisition and amplification platform for sensitive electrochemiluminescent detection of microRNA. Anal Chim Acta 891:130–135CrossRefGoogle Scholar
  31. 31.
    Asadzadeh-Firouzabadi A, Zare HR (2017) Application of cysteamine-capped gold nanoparticles for early detection of lung cancer-specific miRNA (miR-25) in human blood plasma. Anal Methods 9:3852–3861CrossRefGoogle Scholar
  32. 32.
    Asadzadeh-Firouzabadi A, Zare HR (2018) Preparation and application of AgNPs/SWCNTs nanohybrid as an electroactive label for sensitive detection of miRNA related to lung cancer. Sens Actuators B: Chem 260:824–831CrossRefGoogle Scholar
  33. 33.
    Liu H, Bei X, Xia Q, Fu Y, Zhang S, Liu M, Fan K, Zhang M, Yang Y (2016) Enzyme-free electrochemical detection of microRNA-21 using immobilized hairpin probes and a target-triggered hybridization chain reaction amplification strategy. Microchim Acta 183:297–304CrossRefGoogle Scholar
  34. 34.
    Fischer-Posovszky P, Roos J, Kotnik P, Battelino T, Inzaghi E, Nobili V, Cianfarani S, Wabitsch M (2016) Functional significance and predictive value of microRNAs in pediatric obesity: tiny molecules with huge impact? Horm Res Paediatr 86:3–10CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Chemistry, Faculty of ScienceYazd UniversityYazdIran

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