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A new peptide nucleotide acid biosensor for electrochemical detection of single nucleotide polymorphism in duplex DNA via triplex structure formation

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Abstract

In this paper, we report a new PNA biosensor for electrochemical detection of point mutation or single nucleotide polymorphism (SNP) in p53 gene corresponding oligonucleotide based on PNA/ds-DNA triplex formation following hybridization of PNA probe with double-stranded DNA (ds-DNA) sample without denaturing the ds-DNA into single-stranded DNA (ss-DNA). As p53 gene is mutated in many human tumors, this research is useful for cancer therapy and genomic study. In this approach, methylene blue (MB) is used for electrochemical signal generation and the interaction between MB and oligonucleotides is studied by differential pulse voltammety (DPV). Probe-modified electrode is prepared by self-assembled monolayer (SAM) formation of thiolated PNA molecules on the surface of Au electrode. A significant increase in the reduction signal of MB following hybridization of the probe with the complementary double-stranded oligonucleotide (ds-oligonucleotide) confirms the function of the biosensor. The selectivity of the PNA sensor is investigated by non-complementary ds-oligonucleotides and the results support the ability of the sensor to detect single-base mismatch directly on ds-oligonucleotide. The influence of probe and ds-DNA concentrations on the effective discrimination against complementary sequence and point mutation is studied and the concentration of 10−6 M is selected as appropriate concentration. Diagnostic performance of the biosensor is described and the detection limit is found to be 4.15 × 10−12 M.

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Authors and Affiliations

  1. Eletroanalytical Chemistry Research Laboratory, Department of Analytical Chemistry, Faculty of Chemistry, University of Mazandaran, Babolsar, Iran

    Ezat Hamidi-Asl, Jahan Bakhsh Raoof & Reza Ojani

  2. Eletroanalytical Chemistry Research Laboratory, Department of Analytical Chemistry, Faculty of Chemistry, Tabriz University, Tabriz, Iran

    Seyed Mahdi Golabi

  3. Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran

    Mohammad Saeid Hejazi

  4. Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Mohammad Saeid Hejazi

Authors
  1. Ezat Hamidi-Asl
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  2. Jahan Bakhsh Raoof
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  3. Reza Ojani
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  4. Seyed Mahdi Golabi
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  5. Mohammad Saeid Hejazi
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Correspondence to Jahan Bakhsh Raoof.

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Hamidi-Asl, E., Raoof, J.B., Ojani, R. et al. A new peptide nucleotide acid biosensor for electrochemical detection of single nucleotide polymorphism in duplex DNA via triplex structure formation. J IRAN CHEM SOC 10, 1075–1083 (2013). https://doi.org/10.1007/s13738-013-0254-0

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  • DOI: https://doi.org/10.1007/s13738-013-0254-0

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