Indigo Carmine as New Label in PNA Biosensor for Detection of Short Sequence of p53 Tumor Suppressor Gene

Abstract

A new electrochemical PNA hybridization biosensor for detection of a 15-mer sequence unique to p53 using indigo carmine (IC) as an electrochemical detector is described in this work. This genosensor is based on the hybridization of target oligonucleotide with its complementary probe immobilized on the gold electrode by self-assembled monolayer formation. Because this label is electroactive in acidic medium, the interaction between IC and short sequence of p53 is studied by differential pulse voltammety (DPV) in 0.1M H2SO4. The results of electrochemical impedance spectroscopy and cyclic voltammetry in the solution of [Fe(CN)(6)](3-/4-) shows no breakage in PNA-DNA duplex. A decrease in the voltammetric peak currents of IC is observed upon hybridization of the probe with the target DNA. The influence of probe concentration on effective discrimination against non-complementary oligonucleotides is investigated and a concentration of 10(-7)M is selected. The diagnostic performance of the PNA sensor is described and the detection limit is found to be 4.31x10(-12)M.