Label-Free Electrochemical Detection of the Specific Oligonucleotide Sequence of Dengue Virus Type 1 on Pencil Graphite Electrodes

A biosensor that relies on the adsorption immobilization of the 18-mer single-stranded nucleic acid related to dengue virus gene 1 on activated pencil graphite was developed. Hybridization between the probe and its complementary oligonucleotides (the target) was investigated by monitoring guanine oxidation by differential pulse voltammetry (DPV). The pencil graphite electrode was made of ordinary pencil lead (type 4B). The polished surface of the working electrode was activated by applying a potential of 1.8 V for 5 min. Afterward, the dengue oligonucleotides probe was immobilized on the activated electrode by applying 0.5 V to the electrode in 0.5 M acetate buffer (pH 5.0) for 5 min. The hybridization process was carried out by incubating at the annealing temperature of the oligonucleotides. A time of five minutes and concentration of 1 μM were found to be the optimal conditions for probe immobilization. The electrochemical detection of annealing between the DNA probe (TS-1P) immobilized on the modified electrode, and the target (TS-1T) was achieved. The target could be quantified in a range from 1 to 40 nM with good linearity and a detection limit of 0.92 nM. The specificity of the electrochemical biosensor was tested using non-complementary sequences of dengue virus 2 and 3.

[1]  Juan Pablo Tosar,et al.  Two independent label-free detection methods in one electrochemical DNA sensor. , 2009, Biosensors & bioelectronics.

[2]  Marco Mascini,et al.  Electrochemical DNA biosensor for the detection of TT and Hepatitis B virus from PCR amplified real samples by using methylene blue. , 2002, Talanta.

[3]  Melissa M. Eitzen,et al.  Vaccines for Biodefense and Emerging and Neglected Diseases , 2009 .

[4]  Fernando Rodrigues Ribeiro Teles,et al.  Electrochemical Detection of a Dengue-related Oligonucleotide Sequence Using Ferrocenium as a Hybridization Indicator , 2007, Sensors.

[5]  J. Guesdon,et al.  Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue virus serotypes in peripheral blood. , 1990, Journal of virological methods.

[6]  Mohammad Saeid Hejazi,et al.  Brilliant cresyl blue as electroactive indicator in electrochemical DNA oligonucleotide sensors. , 2010, Bioelectrochemistry.

[7]  Bengi Uslu,et al.  Voltammetric studies on the HIV-1 inhibitory drug Efavirenz: the interaction between dsDNA and drug using electrochemical DNA biosensor and adsorptive stripping voltammetric determination on disposable pencil graphite electrode. , 2009, Biosensors & bioelectronics.

[8]  J Wang,et al.  Electrochemical biosensors for DNA hybridization and DNA damage. , 1998, Biosensors & bioelectronics.

[9]  Sibel A. Ozkan,et al.  The Role of and the Place of Method Validation in Drug Analysis Using Electroanalytical Techniques , 2011 .

[10]  M. Hejazi,et al.  Electrochemical detection of human papilloma virus (HPV) target DNA using MB on pencil graphite electrode , 2008 .

[11]  Mehmet Mutlu,et al.  QCM-based DNA biosensor for detection of genetically modified organisms (GMOs) , 2009 .

[12]  Hui Xu,et al.  Electrochemical Sensors for Clinic Analysis , 2008, Sensors.

[13]  Klaus Ammann,et al.  Why farming with high tech methods should integrate elements of organic agriculture. , 2009, New biotechnology.

[14]  N. A. Santana,et al.  Development of electrochemical biosensor for virus dengue diagnostic using graphite electrode , 2009 .

[15]  M. Ozsoz,et al.  Voltammetric studies on the potent carcinogen, 7,12-dimethylbenz[a]anthracene: Adsorptive stripping voltammetric determination in bulk aqueous forms and human urine samples and detection of DNA interaction on pencil graphite electrode. , 2010, Talanta.

[16]  Mohammad Saeid Hejazi,et al.  Differential Pulse Voltammetric Detection of Hepatitis C Virus 1a Oligonucleotide Chain by a Label-Free Electrochemical DNA Hybridization Biosensor Using Consensus Sequence of Hepatitis C Virus 1a Probe on the Pencil Graphite Electrode , 2009 .

[17]  A I Petrov,et al.  Multilayer films containing immobilized nucleic acids. Their structure and possibilities in biosensor applications. , 1996, Biosensors & bioelectronics.

[18]  Da Xing,et al.  PCR-free quantitative detection of genetically modified organism from raw materials. An electrochemiluminescence-based bio bar code method. , 2008, Analytical chemistry.

[19]  Adelair Helena dos Santos,et al.  Dependence of Metarhizium anisopliae on high humidity for ovicidal activity on Aedes aegypti , 2009 .

[20]  M. Guzmán,et al.  Dengue: an update. , 2002, The Lancet. Infectious diseases.

[21]  M. Guzmán,et al.  Dengue diagnosis, advances and challenges. , 2004, International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases.

[22]  Gustavo Rivas,et al.  Indicator-free electrochemical DNA hybridization biosensor , 1998 .

[23]  M. Hejazi,et al.  Developing an electrochemical deoxyribonucleic acid (DNA) biosensor on the basis of human interleukine-2 gene using an electroactive label. , 2006, Analytica chimica acta.

[24]  Mohammad Saeid Hejazi,et al.  Detection of Human Interleukine‐2 Gene Using a Label‐Free Electrochemical DNA Hybridization Biosensor on the Basis of a Non‐Inosine Substituted Probe , 2007 .

[25]  Farrokh Karimi,et al.  Construction, electrochemically biosensing and discrimination of recombinant plasmid (pEThIL-2) on the basis of interleukine-2 DNA insert. , 2008, Biosensors & bioelectronics.

[26]  Kagan Kerman,et al.  Allele-specific genotype detection of factor V Leiden mutation from polymerase chain reaction amplicons based on label-free electrochemical genosensor. , 2002, Analytical chemistry.

[27]  Navid Nasirizadeh,et al.  Introduction of hematoxylin as an electroactive label for DNA biosensors and its employment in detection of target DNA sequence and single-base mismatch in human papilloma virus corresponding to oligonucleotide. , 2011, Biosensors & bioelectronics.

[28]  A. Santana,et al.  Activities of naphthoquinones against Aedes aegypti (Linnaeus, 1762) (Diptera: Culicidae), vector of dengue and Biomphalaria glabrata (Say, 1818), intermediate host of Schistosoma mansoni. , 2009, Acta tropica.

[29]  T. G. Drummond,et al.  Electrochemical DNA sensors , 2003, Nature Biotechnology.

[30]  P. Zanotto,et al.  Phylogeography and evolutionary history of dengue virus type 3. , 2009, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[31]  Duane J. Gubler,et al.  Dengue and Dengue Hemorrhagic Fever , 1998, Clinical Microbiology Reviews.

[32]  Silvia H.P. Serrano,et al.  Electrochemical oxidation of dna , 1995 .

[33]  Richard A Montagna,et al.  Microfluidic biosensor for the serotype-specific detection of dengue virus RNA. , 2005, Analytical chemistry.

[34]  Ezat Hamidi-Asl,et al.  Preparation of an electrochemical PNA biosensor for detection of target DNA sequence and single nucleotide mutation on p53 tumor suppressor gene corresponding oligonucleotide , 2011 .

[35]  M. Domingues,et al.  Inappropriate use of emergency services : a systematic review of prevalence and associated factors , 2009 .

[36]  D. Gubler,et al.  Dengue and dengue hemorrhagic fever. , 2014 .

[37]  Marli Tenório Cordeiro,et al.  Seroprevalence and risk factors for dengue infection in socio-economically distinct areas of Recife, Brazil. , 2010, Acta tropica.

[38]  Yun Kyung Jung,et al.  On-chip colorimetric biosensor based on polydiacetylene (PDA) embedded in photopolymerized poly(ethylene glycol) diacrylate (PEG-DA) hydrogel , 2006 .

[39]  H. S. Wolff,et al.  iRun: Horizontal and Vertical Shape of a Region-Based Graph Compression , 2022, Sensors.

[40]  G. Coelho,et al.  Dengue and Dengue Hemorrhagic Fever, Brazil, 1981–2002 , 2005, Emerging infectious diseases.

[41]  Xiaoru Zhang,et al.  Hybridization biosensor using diaquabis[N-(2-pyridinylmethyl)benzamide-κ2N,O]-cadmium(II) dinitrate as a new electroactive indicator for detection of human hepatitis B virus DNA , 2007 .

[42]  J Wang,et al.  Nucleic-acid immobilization, recognition and detection at chronopotentiometric DNA chips. , 1997, Biosensors & bioelectronics.

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

[44]  M. Barreto,et al.  Dengue: twenty-five years since reemergence in Brazil. , 2009, Cadernos de saude publica.

[45]  A. Baeumner,et al.  RNA biosensor for the rapid detection of viable Escherichia coli in drinking water. , 2003, Biosensors & bioelectronics.