Label-free electrochemical detection of Avian Influenza Virus genotype utilizing multi-walled carbon nanotubes―cobalt phthalocyanine―PAMAM nanocomposite modified glassy carbon electrode

Abstract A novel DNA electrochemical biosensor for label-free determination of DNA sequence related to the Avian Influenza Virus (AIV) genotype was demonstrated in this paper. First, the multi-walled carbon nanotubes–cobalt phthalocyanine (MWNTs–CoPc) nanocomposite and poly (amidoamine) (PAMAM) dendrimer (generation 4.0) were modified on the glassy carbon electrode (GCE) sequentially. Then, DNA probes were successfully immobilized on the modified electrode with G4 PAMAM dendrimer acting as the coupling agent. The hybridization events were monitored by differential pulse voltammetry (DPV) measurement based on the oxidation signals of guanine without any external labels. Under the optimal conditions, the difference in guanine oxidation signal of the probe modified GCE in the absence and presence of complementary target (Δ I p ) was linear with the logarithmic value of the complementary target concentration from 0.01 to 500 ng/ml with a correlation coefficient of 0.998 and a detection limit of 1.0 pg/ml.

[1]  Michael B. Hall,et al.  Dendritic macromolecules: synthesis of starburst dendrimers , 1986 .

[2]  Joseph Wang,et al.  Carbon-nanotube-modified glassy carbon electrodes for amplified label-free electrochemical detection of DNA hybridization. , 2003, The Analyst.

[3]  M. Peiris,et al.  Clinical features and rapid viral diagnosis of human disease associated with avian influenza A H5N1 virus , 1998, The Lancet.

[4]  A. Abbaspour,et al.  Electrocatalytic oxidation of guanine and ss-DNA at a cobalt (II) phthalocyanine modified carbon paste electrode , 2004 .

[5]  Changgui Li,et al.  Real-time RT-PCR for H5N1 avian influenza A virus detection. , 2007, Journal of medical microbiology.

[6]  Guo-Li Shen,et al.  Functionalization of multi-walled carbon nanotubes with poly(amidoamine) dendrimer for mediator-free glucose biosensor , 2007 .

[7]  T. Nyokong,et al.  Electrocatalysis of asulam on cobalt phthalocyanine modified multi-walled carbon nanotubes immobilized on a basal plane pyrolytic graphite electrode , 2006 .

[8]  Q. Ma,et al.  A nitrite biosensor based on the immobilization of cytochrome c on multi-walled carbon nanotubes-PAMAM-chitosan nanocomposite modified glass carbon electrode. , 2009, Biosensors & bioelectronics.

[9]  Tao Yang,et al.  A DNA electrochemical sensor based on nanogold-modified poly-2,6-pyridinedicarboxylic acid film and detection of PAT gene fragment. , 2007, Analytical biochemistry.

[10]  P. Eklund,et al.  Debundling and dissolution of single-walled carbon nanotubes in amide solvents. , 2004, Journal of the American Chemical Society.

[11]  Akikazu Sakudo,et al.  Efficient capture of infectious H5 avian influenza virus utilizing magnetic beads coated with anionic polymer. , 2008, Biochemical and biophysical research communications.

[12]  D. Ozkan,et al.  Label‐Free Bioelectronic Detection of Point Mutation by Using Peptide Nucleic Acid Probes , 2003 .

[13]  Mehmet Ozsoz,et al.  Label-free electrochemical hybridization genosensor for the detection of hepatitis B virus genotype on the development of Lamivudine resistance. , 2005, Analytical chemistry.

[14]  Kotaro Suzuki,et al.  Subtyping of Avian Influenza Viruses H1 to H15 on the Basis of Hemagglutinin Genes by PCR Assay and Molecular Determination of Pathogenic Potential , 2008, Journal of Clinical Microbiology.

[15]  Alan Mathewson,et al.  Investigation into the effect that probe immobilisation method type has on the analytical signal of an EIS DNA biosensor. , 2007, Biosensors & bioelectronics.

[16]  Guonan Chen,et al.  Electrochemical biosensor based on hairpin DNA probe using 2-nitroacridone as electrochemical indicator for detection of DNA species related to Chronic Myelogenous Leukemia , 2008 .

[17]  T. Nyokong,et al.  Electrooxidation of hydrazine catalyzed by noncovalently functionalized single-walled carbon nanotubes with CoPc , 2008 .

[18]  Pedro Estrela,et al.  Label-free electrical detection of DNA hybridization for the example of influenza virus gene sequences. , 2008, Analytical biochemistry.

[19]  Guo Hongxia,et al.  Electrochemical detection of short sequences related to the hepatitis B virus using MB on chitosan-modified CPE. , 2007, Bioelectrochemistry.

[20]  Guozhong Zhang,et al.  Inhibition of highly pathogenic avian H5N1 influenza virus replication by RNA oligonucleotides targeting NS1 gene. , 2008, Biochemical and biophysical research communications.

[21]  A. Erdem,et al.  Rigid carbon composites: a new transducing material for label-free electrochemical genosensing , 2004 .

[22]  Y. Guan,et al.  Performance Evaluation of Five Detection Tests for Avian Influenza Antigen with Various Avian Samples , 2007, Avian diseases.