Electrochemical detection of DNA hybridization using a water-soluble branched polyethyleneimine–cobalt(III)–phenanthroline indicator and PNA probe on Au electrodes

Abstract

An electrochemical method for the detection of DNA hybridization using a novel electroactive, cationic, and water-soluble branched polyethyleneimine (BPEI)–cobalt(III)–phenanthroline(phen) polymeric indicator and single-stranded neutral peptide nucleic acid (PNA) probe on the Au electrode was developed. The indicator possesses some free amine groups, as well as cationic cobalt complexes in the polymer chain. It does not bind to neutral PNA capture probe alone. However, the indicator strongly interacts with the negatively charged backbone of the complementary oligonucleotide bound to the PNA probe through electrostatic interactions. The coordination spherical moieties also interact with the probe by embedding into the double-helix structure of PNA–DNA. These two interactions enable transduction of hybridization, producing a clear electrical signal in differential pulse voltammetry (DPV). The correlation against non-complementary DNA, three-base and one-base mismatch DNA was sharp, and the signal of indicator for the target DNA demonstrated a linear relationship within the concentration range of 5.0 × 10 −9 to 2.5 × 10 −7 M ( R = 0.9940) with a detection limit of 5.6 × 10 −10 M. These studies showed that the novel polymeric indicator and single-stranded PNA probe could be used to fabricate an electrochemical biosensor for DNA detection. This technique can provide an alternative route for expanding the range of detection methods available for DNA hybridization.