A new biosensing strategy for direct electrochemical detection of DNA hybridization by AC impedance measurement is described. The indicator or label free approach was developed on the basis of the carbon nanotubes coupled with a conductive polymer polypyrrole (PPy). Multi-walled carbon nanotubes functionalized with carboxylic group (MWNTs-COOH) were modified on the glassy carbon electrode (GCE) and the oligonucleotide probes were doped within the electro-polymerized PPy films by serving as the sole counter anion during the growth of the conducting films. Before and after hybridization reaction with the complementary DNA sequences, a decrease of impedance values was observed as a result of the reduction of the electrode resistance. Hybridization amounts of the one-base, two-base and three-base mismatched sequences were obtained only in a 51%, 18% and 8.2% response when compared to that for the complementary matched sequence. Such unique response is attributed to the concomitant conductivity changes of the PPy-polymerized carbon nanotubes, and offers great promise for reagentless DNA hybridization analysis.
The electrochemical detection of sequence-specific DNA using a DNA probe labeled with aminoferrocene (AFC) is reported. Sample ssDNA was immobilized on a chitosan modified glassy carbon electrode. A sequence-known DNA with 256 bp [obtained by polymerase chain reaction (PCR)] was successfully labeled with the electro-active reagent AFC by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide for the first time. This DNA probe labeled with AFC was applied to hybridize with a sequence-unknown DNA sample. Only the complementary sequence (cDNA) could form a double-stranded DNA (dsDNA) with the DNA probe labeled with AFC. The anodic peak currents (ipa) of the AFC bound to the dsDNA by differential pulse voltammetry were used for the determination of cDNA. The ipa of AFC was linearly related to the concentration of cDNA sequence between 1.0 x 10(-8) and 6.0 x 10(-6) mol L-1. The detection limit was 2.0 x 10(-9) mol L-1 using 3 sigma (where sigma is the standard deviation of blank solution, n = 11). The probe showed high sensitivity and selectivity.
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