Electronically conductive polymer grafted with oligonucleotides as electrosensors of DNA: Preliminary study of real time monitoring by in situ techniques

Polypyrrole bearing covalently linked oligonucleotide (ODN) probes has been used for electrode modification through one-step electrosynthesis. The biofilms obtained were employed for the direct and real-time detection of DNA hybridisation by using photocurrent spectroscopy and a quartz crystal microbalance (QCM). The photoelectrochemical properties of polypyrrole appear to be influenced by the presence of ODN allowing the transduction of the hybridisation event into a measurable photocurrent. The operational parameters of the biosensor (film thickness, ODN load, …) were assessed to optimise the response to DNA hybridisation. In addition, QCM detection, which is based on the monitoring of the biofilm mass uptake generated by molecular recognition, has been used as a validation technique. The sensors obtained have been demonstrated to be fully reusable. They have also shown a suitable sensitivity and a good reproducibility in terms of kinetic behaviour and intensity of the measured signal. Finally, this study considers the pertinence of both techniques regarding DNA hybridisation detection.

[1]  Hafsa Korri-Youssoufi,et al.  Toward Bioelectronics: Specific DNA Recognition Based on an Oligonucleotide-Functionalized Polypyrrole , 1997 .

[2]  L. Abrantes,et al.  Study of DNA hybridization on polypyrrole grafted with oligonucleotides by photocurrent spectroscopy. , 2001, Biosensors & bioelectronics.

[3]  M. Thompson,et al.  Hybridization of complementary strand and single-base mutated oligonucleotides detected with an on-line acoustic wave sensor. , 1998, The Analyst.

[4]  B. D. Malhotra,et al.  Recent studies of heterocyclic and aromatic conducting polymers , 1986 .

[5]  Constantin Polychronakos,et al.  Functionalization of Si/SiO2 substrates with homooligonucleotides for a DNA biosensor , 1999 .

[6]  Joseph Wang,et al.  New label-free DNA recognition based on doping nucleic-acid probes within conducting polymer films , 1999 .

[7]  H M Widmer,et al.  Fiber-optic evanescent wave biosensor for the detection of oligonucleotides. , 1996, Analytical chemistry.

[8]  L. Abrantes,et al.  Photoelectrochemical studies of polymer films: Poly(3-methylthiophene) and poly(3-methylthiophene)/Cu systems , 1996 .

[9]  G L Hayward,et al.  Acoustic waves and the study of biochemical macromolecules and cells at the sensor-liquid interface. , 1999, The Analyst.

[10]  Y. Okahata,et al.  Kinetic measurements of DNA hybridization on an oligonucleotide-immobilized 27-MHz quartz crystal microbalance. , 1998, Analytical chemistry.

[11]  Yoshio Okahata,et al.  Hybridization of nucleic acids immobilized on a quartz crystal microbalance , 1992 .

[12]  D. Guyomard.  Mise au Principes de base de l'électrochimie des semi-conducteurs , 1986 .

[13]  Wolfgang W. Gärtner,et al.  Depletion-Layer Photoeffects in Semiconductors , 1959 .

[14]  Stanley Bruckenstein,et al.  Experimental aspects of use of the quartz crystal microbalance in solution , 1985 .

[15]  Michael B. Thompson,et al.  Interfacial Nucleic Acid Hybridization Studied by Random Primer 32P Labeling and Liquid-Phase Acoustic Network Analysis , 1994 .

[16]  Christina G. Siontorou,et al.  Rapid methods for detection of Aflatoxin M1 based on electrochemical transduction by self-assembled metal-supported bilayer lipid membranes (s-BLMs) and on interferences with transduction of DNA hybridization , 1998 .

[17]  John F. Vetelino,et al.  An acoustic plate mode biosensor , 1992 .

[18]  Giovanna Marrazza,et al.  Disposable DNA electrochemical biosensors for environmental monitoring , 1999 .

[19]  P. Nilsson,et al.  Analysis of oligonucleotide probe affinities using surface plasmon resonance: a means for mutational scanning. , 1997, Analytical biochemistry.

[20]  D. Allara,et al.  Real Time Spectroscopic Ellipsometry: In Situ Characterization of Pyrrole Electropolymerization , 1991 .

[21]  M. McGovern,et al.  Surface immobilized biochemical macromolecules studied by scanning Kelvin microprobe. , 2000, Faraday discussions.

[22]  T. Livache,et al.  Polypyrrole DNA chip on a silicon device: example of hepatitis C virus genotyping. , 1998, Analytical biochemistry.

[23]  G. Bidan,et al.  Spectroelectrochemical study of polypyrrole films , 1983 .

[24]  Joseph Wang,et al.  Towards Genoelectronics: Electrochemical Biosensing of DNA Hybridization , 1999 .

[25]  G. Dryhurst,et al.  Electrochemical Oxidation of Guanine at the Pyrolytic Graphite Electrode , 1970 .

[26]  John Alderman,et al.  A DNA diagnostic biosensor: development, characterisation and performance , 2000 .

[27]  F Lesbre,et al.  Characterization and optimization of a real-time, parallel, label-free, polypyrrole-based DNA sensor by surface plasmon resonance imaging. , 2000, Analytical chemistry.

[28]  T. Livache,et al.  Preparation of a DNA matrix via an electrochemically directed copolymerization of pyrrole and oligonucleotides bearing a pyrrole group. , 1994, Nucleic acids research.