title: Electrochemical detection of the MT-ND6 gene and its enzymatic digestion: application in human genomic sample.
abstract: A simple electrochemical biosensor was developed for the detection of the mitochondrial NADH dehydrogenase 6 gene (MT-ND6) and its enzymatic digestion by BamHI enzyme. This biosensor was fabricated by modification of a glassy carbon electrode with gold nanoparticles (AuNPs/GCE) and a probe oligonucleotide (ssDNA/AuNPs/GCE). The probe, which is a thiolated segment of the MT-ND6 gene, was deposited by self-assembling immobilization on AuNPs/GCE. Two indicators including methylene blue (MB) and neutral red (NR) were used as the electroactive indicators and the electrochemical response of the modified electrode was measured by differential pulse voltammetry. The proposed biosensor can detect the complementary sequences of the MT-ND6 gene. Also the modified electrode was used for the detection of an enzymatic digestion process by BamHI enzyme. The electrochemical biosensor can detect the MT-ND6 gene and its enzymatic digestion in polymerase chain reaction (PCR)-amplified DNA extracted from human blood. Also the biosensor was used directly for detection of the MT-ND6 gene in all of the human genome.
authors.affiliation: Department of Chemistry, Faculty of Science, Yazd University, Yazd, Islamic Republic of Iran. Electronic address: mazloum@yazduni.ac.ir.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Mazloum-Ardakani
authors.affiliation: Department of Chemistry, Faculty of Science, Yazd University, Yazd, Islamic Republic of Iran.
authors.forename: Roya
authors.initials: R
authors.lastname: Ahmadi
authors.affiliation: Department of Biology, Faculty of Science, Yazd University, Yazd, Islamic Republic of Iran.
authors.forename: Mohammad Mehdi
authors.initials: MM
authors.lastname: Heidari
authors.affiliation: Shahid Bakeri High Education Center of Miandoab, Urmia University, Urmia, Islamic Republic of Iran.
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Sheikh-Mohseni
mesh_terms: D015374:Biosensing Techniques; D015245:Deoxyribonuclease BamHI / Q000378:metabolism; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D015894:Genome, Human; D006046:Gold; D006801:Humans; D009245:NADH Dehydrogenase / Q000032:analysis* / Q000097:blood / Q000378:metabolism; D053758:Nanoparticles; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D009841:Oligonucleotides; D006046:Gold; C507225:MT-ND6 protein, human; D009245:NADH Dehydrogenase; D015245:Deoxyribonuclease BamHI
keywords: DNA hybridization; Electrochemical biosensor; Enzyme digestion; Genome sample; Gold nanoparticles