Direct detection and discrimination of double-stranded oligonucleotide corresponding to hepatitis C virus genotype 3a using an electrochemical DNA biosensor based on peptide nucleic acid and double-stranded DNA hybridization
Development of an electrochemical DNA biosensor for the direct detection and discrimination of double-stranded oligonucleotide (dsDNA) corresponding to hepatitis C virus genotype 3a, without its denaturation, using a gold electrode is described. The electrochemical DNA sensor relies on the modificat...
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| Veröffentlicht in: | Analytical and bioanalytical chemistry 2010-08, Vol.397 (8), p.3581-3587 |
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| creator | Pournaghi-Azar, M. H Ahour, F Hejazi, M. S |
| description | Development of an electrochemical DNA biosensor for the direct detection and discrimination of double-stranded oligonucleotide (dsDNA) corresponding to hepatitis C virus genotype 3a, without its denaturation, using a gold electrode is described. The electrochemical DNA sensor relies on the modification of the gold electrode with 6-mercapto-1-hexanol and a self-assembled monolayer of 14-mer peptide nucleic acid probe, related to the hepatitis C virus genotype 3a core/E1 region. The increase of differential pulse voltammetric responses of methylene blue, upon hybridization of the self-assembled probe with the target ds-DNA to form a triplex is the principle behind the detection and discrimination. Some hybridization experiments with non-complementary oligonucleotides were carried out to assess whether the developed DNA sensor responds selectively to the ds-DNA target. Diagnostic performance of the biosensor is described and the detection limit was found to be 1.8 × 10⁻¹² M in phosphate buffer solution, pH 7.0. The relative standard deviation of measurements of 100 pM of target ds-DNA performed with three independent probe-modified electrodes was 3.1%, indicating a remarkable reproducibility of the detection method. [graphic removed] |
| doi_str_mv | 10.1007/s00216-010-3875-5 |
| format | Article |
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H ; Ahour, F ; Hejazi, M. S</creator><creatorcontrib>Pournaghi-Azar, M. H ; Ahour, F ; Hejazi, M. S</creatorcontrib><description>Development of an electrochemical DNA biosensor for the direct detection and discrimination of double-stranded oligonucleotide (dsDNA) corresponding to hepatitis C virus genotype 3a, without its denaturation, using a gold electrode is described. The electrochemical DNA sensor relies on the modification of the gold electrode with 6-mercapto-1-hexanol and a self-assembled monolayer of 14-mer peptide nucleic acid probe, related to the hepatitis C virus genotype 3a core/E1 region. The increase of differential pulse voltammetric responses of methylene blue, upon hybridization of the self-assembled probe with the target ds-DNA to form a triplex is the principle behind the detection and discrimination. Some hybridization experiments with non-complementary oligonucleotides were carried out to assess whether the developed DNA sensor responds selectively to the ds-DNA target. Diagnostic performance of the biosensor is described and the detection limit was found to be 1.8 × 10⁻¹² M in phosphate buffer solution, pH 7.0. The relative standard deviation of measurements of 100 pM of target ds-DNA performed with three independent probe-modified electrodes was 3.1%, indicating a remarkable reproducibility of the detection method. [graphic removed]</description><identifier>ISSN: 1618-2642</identifier><identifier>EISSN: 1618-2650</identifier><identifier>DOI: 10.1007/s00216-010-3875-5</identifier><identifier>PMID: 20563795</identifier><language>eng</language><publisher>Berlin/Heidelberg: Berlin/Heidelberg : Springer-Verlag</publisher><subject>Analysis ; Analytical Chemistry ; Biochemistry ; Biological and medical sciences ; Biosensing Techniques - instrumentation ; Biosensing Techniques - methods ; Biosensors ; Biotechnology ; Characterization and Evaluation of Materials ; Chemical properties ; Chemistry ; Chemistry and Materials Science ; Deoxyribonucleic acid ; Design and construction ; Discrimination ; DNA biosensor ; DNA, Viral - analysis ; DNA, Viral - genetics ; Electrochemical analysis ; Electrochemical methods ; Electrodes ; Exact sciences and technology ; Food Science ; Fundamental and applied biological sciences. Psychology ; General, instrumentation ; Genetic aspects ; Genotype ; Gold ; Gold electrode ; Hepacivirus - genetics ; Hepatitis C virus ; Laboratory Medicine ; Limit of Detection ; Methods ; Methods. Procedures. 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H</creatorcontrib><creatorcontrib>Ahour, F</creatorcontrib><creatorcontrib>Hejazi, M. S</creatorcontrib><title>Direct detection and discrimination of double-stranded oligonucleotide corresponding to hepatitis C virus genotype 3a using an electrochemical DNA biosensor based on peptide nucleic acid and double-stranded DNA hybridization</title><title>Analytical and bioanalytical chemistry</title><addtitle>Anal Bioanal Chem</addtitle><addtitle>Anal Bioanal Chem</addtitle><description>Development of an electrochemical DNA biosensor for the direct detection and discrimination of double-stranded oligonucleotide (dsDNA) corresponding to hepatitis C virus genotype 3a, without its denaturation, using a gold electrode is described. The electrochemical DNA sensor relies on the modification of the gold electrode with 6-mercapto-1-hexanol and a self-assembled monolayer of 14-mer peptide nucleic acid probe, related to the hepatitis C virus genotype 3a core/E1 region. The increase of differential pulse voltammetric responses of methylene blue, upon hybridization of the self-assembled probe with the target ds-DNA to form a triplex is the principle behind the detection and discrimination. Some hybridization experiments with non-complementary oligonucleotides were carried out to assess whether the developed DNA sensor responds selectively to the ds-DNA target. Diagnostic performance of the biosensor is described and the detection limit was found to be 1.8 × 10⁻¹² M in phosphate buffer solution, pH 7.0. The relative standard deviation of measurements of 100 pM of target ds-DNA performed with three independent probe-modified electrodes was 3.1%, indicating a remarkable reproducibility of the detection method. [graphic removed]</description><subject>Analysis</subject><subject>Analytical Chemistry</subject><subject>Biochemistry</subject><subject>Biological and medical sciences</subject><subject>Biosensing Techniques - instrumentation</subject><subject>Biosensing Techniques - methods</subject><subject>Biosensors</subject><subject>Biotechnology</subject><subject>Characterization and Evaluation of Materials</subject><subject>Chemical properties</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Deoxyribonucleic acid</subject><subject>Design and construction</subject><subject>Discrimination</subject><subject>DNA biosensor</subject><subject>DNA, Viral - analysis</subject><subject>DNA, Viral - genetics</subject><subject>Electrochemical analysis</subject><subject>Electrochemical methods</subject><subject>Electrodes</subject><subject>Exact sciences and technology</subject><subject>Food Science</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>General, instrumentation</subject><subject>Genetic aspects</subject><subject>Genotype</subject><subject>Gold</subject><subject>Gold electrode</subject><subject>Hepacivirus - genetics</subject><subject>Hepatitis C virus</subject><subject>Laboratory Medicine</subject><subject>Limit of Detection</subject><subject>Methods</subject><subject>Methods. Procedures. Technologies</subject><subject>Monitoring/Environmental Analysis</subject><subject>Monomolecular films</subject><subject>Nucleic Acid Conformation</subject><subject>Nucleic Acid Hybridization</subject><subject>Nucleic acid probes</subject><subject>Nucleic acids</subject><subject>Oligonucleotides</subject><subject>Oligonucleotides - chemistry</subject><subject>Oligonucleotides - genetics</subject><subject>Original Paper</subject><subject>Peptide Nucleic Acids - analysis</subject><subject>Peptide Nucleic Acids - genetics</subject><subject>Peptides</subject><subject>PNA probe</subject><subject>Triplex formation</subject><subject>Various methods and equipments</subject><subject>viral hepatitis</subject><issn>1618-2642</issn><issn>1618-2650</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFks1u1TAQhSMEoqXwAGzAUoVgk-KfxHGWVy1_UgUL6Npy7Mmtq1w72AlSeVoehclNKQKhoiwc2d-ZM5o5RfGU0RNGafM6U8qZLCmjpVBNXdb3ikMmmSq5rOn92_-KHxSPcr6ilNWKyYfFAae1FE1bHxY_znwCOxEHEx4-BmKCI85nm_zOB7O_ij1xce4GKPOU8B0ciYPfxjDbAeLkHRAbU4I8xuB82JIpkksYUTz5TE7JN5_mTLYQ4nQ9AhGGzHnBTCAwoG2K9hJ23pqBnH3ckM7HDCHHRDqTF69ARhj3NntHb4mx3q2d_tXXor-87pJ3_vu--cfFg94MGZ7cnEfFxds3X07fl-ef3n043ZyXtubtVCrR9co1ktpeCWetoB0oDrLvKl5VyrR95zjj1CpmlFV1R9u2ajkyTaVwDeKoeLnWHVP8OkOe9A6HCMNgAsQ5ayVbJRir1H9JLIiobBby1Z0kkw0TbVMziejxim7NANqHPuJE7ILrjWgVawVF9qg4-QeFn1vGHwP0Hu__ELBVYFPMOUGvR8yFSdeaUb1kUK8Z1JhBvWRQ16h5dtP13O3A3Sp-hQ6BFzeAybjzHldnff7NCdpyISvk-MplfApbSPoqzingFu90f76KehO12SYsfPGZUyYoU42QWPYnqTEAHg</recordid><startdate>20100801</startdate><enddate>20100801</enddate><creator>Pournaghi-Azar, M. 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Psychology</topic><topic>General, instrumentation</topic><topic>Genetic aspects</topic><topic>Genotype</topic><topic>Gold</topic><topic>Gold electrode</topic><topic>Hepacivirus - genetics</topic><topic>Hepatitis C virus</topic><topic>Laboratory Medicine</topic><topic>Limit of Detection</topic><topic>Methods</topic><topic>Methods. Procedures. Technologies</topic><topic>Monitoring/Environmental Analysis</topic><topic>Monomolecular films</topic><topic>Nucleic Acid Conformation</topic><topic>Nucleic Acid Hybridization</topic><topic>Nucleic acid probes</topic><topic>Nucleic acids</topic><topic>Oligonucleotides</topic><topic>Oligonucleotides - chemistry</topic><topic>Oligonucleotides - genetics</topic><topic>Original Paper</topic><topic>Peptide Nucleic Acids - analysis</topic><topic>Peptide Nucleic Acids - genetics</topic><topic>Peptides</topic><topic>PNA probe</topic><topic>Triplex formation</topic><topic>Various methods and equipments</topic><topic>viral hepatitis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pournaghi-Azar, M. H</creatorcontrib><creatorcontrib>Ahour, F</creatorcontrib><creatorcontrib>Hejazi, M. 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H</au><au>Ahour, F</au><au>Hejazi, M. S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Direct detection and discrimination of double-stranded oligonucleotide corresponding to hepatitis C virus genotype 3a using an electrochemical DNA biosensor based on peptide nucleic acid and double-stranded DNA hybridization</atitle><jtitle>Analytical and bioanalytical chemistry</jtitle><stitle>Anal Bioanal Chem</stitle><addtitle>Anal Bioanal Chem</addtitle><date>2010-08-01</date><risdate>2010</risdate><volume>397</volume><issue>8</issue><spage>3581</spage><epage>3587</epage><pages>3581-3587</pages><issn>1618-2642</issn><eissn>1618-2650</eissn><abstract>Development of an electrochemical DNA biosensor for the direct detection and discrimination of double-stranded oligonucleotide (dsDNA) corresponding to hepatitis C virus genotype 3a, without its denaturation, using a gold electrode is described. The electrochemical DNA sensor relies on the modification of the gold electrode with 6-mercapto-1-hexanol and a self-assembled monolayer of 14-mer peptide nucleic acid probe, related to the hepatitis C virus genotype 3a core/E1 region. The increase of differential pulse voltammetric responses of methylene blue, upon hybridization of the self-assembled probe with the target ds-DNA to form a triplex is the principle behind the detection and discrimination. Some hybridization experiments with non-complementary oligonucleotides were carried out to assess whether the developed DNA sensor responds selectively to the ds-DNA target. Diagnostic performance of the biosensor is described and the detection limit was found to be 1.8 × 10⁻¹² M in phosphate buffer solution, pH 7.0. The relative standard deviation of measurements of 100 pM of target ds-DNA performed with three independent probe-modified electrodes was 3.1%, indicating a remarkable reproducibility of the detection method. [graphic removed]</abstract><cop>Berlin/Heidelberg</cop><pub>Berlin/Heidelberg : Springer-Verlag</pub><pmid>20563795</pmid><doi>10.1007/s00216-010-3875-5</doi><tpages>7</tpages></addata></record> |
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| ispartof | Analytical and bioanalytical chemistry, 2010-08, Vol.397 (8), p.3581-3587 |
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| subjects | Analysis Analytical Chemistry Biochemistry Biological and medical sciences Biosensing Techniques - instrumentation Biosensing Techniques - methods Biosensors Biotechnology Characterization and Evaluation of Materials Chemical properties Chemistry Chemistry and Materials Science Deoxyribonucleic acid Design and construction Discrimination DNA biosensor DNA, Viral - analysis DNA, Viral - genetics Electrochemical analysis Electrochemical methods Electrodes Exact sciences and technology Food Science Fundamental and applied biological sciences. Psychology General, instrumentation Genetic aspects Genotype Gold Gold electrode Hepacivirus - genetics Hepatitis C virus Laboratory Medicine Limit of Detection Methods Methods. Procedures. Technologies Monitoring/Environmental Analysis Monomolecular films Nucleic Acid Conformation Nucleic Acid Hybridization Nucleic acid probes Nucleic acids Oligonucleotides Oligonucleotides - chemistry Oligonucleotides - genetics Original Paper Peptide Nucleic Acids - analysis Peptide Nucleic Acids - genetics Peptides PNA probe Triplex formation Various methods and equipments viral hepatitis |
| title | Direct detection and discrimination of double-stranded oligonucleotide corresponding to hepatitis C virus genotype 3a using an electrochemical DNA biosensor based on peptide nucleic acid and double-stranded DNA hybridization |
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