Immobilization-free sequence-specific electrochemical detection of DNA using ferrocene-labeled peptide nucleic acid.
Identifieur interne : 002299 ( Main/Exploration ); précédent : 002298; suivant : 002300Immobilization-free sequence-specific electrochemical detection of DNA using ferrocene-labeled peptide nucleic acid.
Auteurs : RBID : pubmed:18771276English descriptors
- KwdEn :
- Base Pair Mismatch, Base Sequence, DNA, Complementary (analysis), DNA, Complementary (genetics), Electrochemical Techniques (methods), Electrodes, Ferrous Compounds (chemistry), Peptide Nucleic Acids (chemistry), Polyamines (chemistry), Polymerase Chain Reaction (methods), Substrate Specificity, Tin Compounds (chemistry).
- MESH :
- chemical , analysis : DNA, Complementary.
- chemical , chemistry : Ferrous Compounds, Peptide Nucleic Acids, Polyamines, Tin Compounds.
- chemical , genetics : DNA, Complementary.
- methods : Electrochemical Techniques, Polymerase Chain Reaction.
- Base Pair Mismatch, Base Sequence, Electrodes, Substrate Specificity.
Abstract
An electrochemical method for sequence-specific detection of DNA without solid-phase probe immobilization is reported. This detection scheme starts with a solution-phase hybridization of ferrocene-labeled peptide nucleic acid (Fc-PNA) and its complementary DNA (cDNA) sequence, followed by the electrochemical transduction of Fc-PNA-DNA hybrid on indium tin oxide (ITO)-based substrates. On the bare ITO electrode, the negatively charged Fc-PNA-DNA hybrid exhibits a much reduced electrochemical signal than that of the neutral-charge Fc-PNA. This is attributed to the electrostatic repulsion between the negatively charged ITO surface and the negatively charged DNA, hindering the access of Fc-PNA-DNA to the electrode. On the contrary, when the transduction measurement is done on the ITO electrode coated with a positively charged poly(allylamine hydrochloride) (PAH) layer, the electrostatic attraction between the (+) PAH surface and the (-) Fc-PNA-DNA hybrid leads to a much higher electrochemical signal than that of the Fc-PNA. The measured electrochemical signal is proportional to the amount of cDNA present. In terms of detection sensitivity, the PAH-modified ITO platform was found to be more sensitive (with a detection limit of 40 fmol) than the bare ITO counterpart (with a detection limit of 500 fmol). At elevated temperatures, this method was able to distinguish fully matched target DNA from DNA with partial mismatches. Unpurified PCR amplicons were detected using a similar format with a detection limit down to 4.17 amol. This detection method holds great promise for single-base mismatch detection as well as electrochemistry-based detection of post-PCR products.
DOI: 10.1021/ac8010236
PubMed: 18771276
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Immobilization-free sequence-specific electrochemical detection of DNA using ferrocene-labeled peptide nucleic acid.</title><author><name sortKey="Luo, Xiaoteng" uniqKey="Luo X">Xiaoteng Luo</name><affiliation wicri:level="1"><nlm:affiliation>Bioengineering Graduate Program, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.</nlm:affiliation><country xml:lang="fr">Hong Kong</country><wicri:regionArea>Bioengineering Graduate Program, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon</wicri:regionArea></affiliation></author><author><name sortKey="Lee, Thomas Ming Hung" uniqKey="Lee T">Thomas Ming-Hung Lee</name></author><author><name sortKey="Hsing, I Ming" uniqKey="Hsing I">I-Ming Hsing</name></author></titleStmt><publicationStmt><date when="2008">2008</date><idno type="doi">10.1021/ac8010236</idno><idno type="RBID">pubmed:18771276</idno><idno type="pmid">18771276</idno><idno type="wicri:Area/Main/Corpus">002247</idno><idno type="wicri:Area/Main/Curation">002247</idno><idno type="wicri:Area/Main/Exploration">002299</idno></publicationStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Pair Mismatch</term><term>Base Sequence</term><term>DNA, Complementary (analysis)</term><term>DNA, Complementary (genetics)</term><term>Electrochemical Techniques (methods)</term><term>Electrodes</term><term>Ferrous Compounds (chemistry)</term><term>Peptide Nucleic Acids (chemistry)</term><term>Polyamines (chemistry)</term><term>Polymerase Chain Reaction (methods)</term><term>Substrate Specificity</term><term>Tin Compounds (chemistry)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>DNA, Complementary</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Ferrous Compounds</term><term>Peptide Nucleic Acids</term><term>Polyamines</term><term>Tin Compounds</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Complementary</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Electrochemical Techniques</term><term>Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Pair Mismatch</term><term>Base Sequence</term><term>Electrodes</term><term>Substrate Specificity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">An electrochemical method for sequence-specific detection of DNA without solid-phase probe immobilization is reported. This detection scheme starts with a solution-phase hybridization of ferrocene-labeled peptide nucleic acid (Fc-PNA) and its complementary DNA (cDNA) sequence, followed by the electrochemical transduction of Fc-PNA-DNA hybrid on indium tin oxide (ITO)-based substrates. On the bare ITO electrode, the negatively charged Fc-PNA-DNA hybrid exhibits a much reduced electrochemical signal than that of the neutral-charge Fc-PNA. This is attributed to the electrostatic repulsion between the negatively charged ITO surface and the negatively charged DNA, hindering the access of Fc-PNA-DNA to the electrode. On the contrary, when the transduction measurement is done on the ITO electrode coated with a positively charged poly(allylamine hydrochloride) (PAH) layer, the electrostatic attraction between the (+) PAH surface and the (-) Fc-PNA-DNA hybrid leads to a much higher electrochemical signal than that of the Fc-PNA. The measured electrochemical signal is proportional to the amount of cDNA present. In terms of detection sensitivity, the PAH-modified ITO platform was found to be more sensitive (with a detection limit of 40 fmol) than the bare ITO counterpart (with a detection limit of 500 fmol). At elevated temperatures, this method was able to distinguish fully matched target DNA from DNA with partial mismatches. Unpurified PCR amplicons were detected using a similar format with a detection limit down to 4.17 amol. This detection method holds great promise for single-base mismatch detection as well as electrochemistry-based detection of post-PCR products.</div></front></TEI><pubmed><MedlineCitation Owner="NLM" Status="MEDLINE"><PMID Version="1">18771276</PMID><DateCreated><Year>2008</Year><Month>10</Month><Day>02</Day></DateCreated><DateCompleted><Year>2008</Year><Month>11</Month><Day>25</Day></DateCompleted><DateRevised><Year>2012</Year><Month>11</Month><Day>15</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1520-6882</ISSN><JournalIssue CitedMedium="Internet"><Volume>80</Volume><Issue>19</Issue><PubDate><Year>2008</Year><Month>Oct</Month><Day>1</Day></PubDate></JournalIssue><Title>Analytical chemistry</Title><ISOAbbreviation>Anal. Chem.</ISOAbbreviation></Journal><ArticleTitle>Immobilization-free sequence-specific electrochemical detection of DNA using ferrocene-labeled peptide nucleic acid.</ArticleTitle><Pagination><MedlinePgn>7341-6</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1021/ac8010236</ELocationID><Abstract><AbstractText>An electrochemical method for sequence-specific detection of DNA without solid-phase probe immobilization is reported. This detection scheme starts with a solution-phase hybridization of ferrocene-labeled peptide nucleic acid (Fc-PNA) and its complementary DNA (cDNA) sequence, followed by the electrochemical transduction of Fc-PNA-DNA hybrid on indium tin oxide (ITO)-based substrates. On the bare ITO electrode, the negatively charged Fc-PNA-DNA hybrid exhibits a much reduced electrochemical signal than that of the neutral-charge Fc-PNA. This is attributed to the electrostatic repulsion between the negatively charged ITO surface and the negatively charged DNA, hindering the access of Fc-PNA-DNA to the electrode. On the contrary, when the transduction measurement is done on the ITO electrode coated with a positively charged poly(allylamine hydrochloride) (PAH) layer, the electrostatic attraction between the (+) PAH surface and the (-) Fc-PNA-DNA hybrid leads to a much higher electrochemical signal than that of the Fc-PNA. The measured electrochemical signal is proportional to the amount of cDNA present. In terms of detection sensitivity, the PAH-modified ITO platform was found to be more sensitive (with a detection limit of 40 fmol) than the bare ITO counterpart (with a detection limit of 500 fmol). At elevated temperatures, this method was able to distinguish fully matched target DNA from DNA with partial mismatches. Unpurified PCR amplicons were detected using a similar format with a detection limit down to 4.17 amol. This detection method holds great promise for single-base mismatch detection as well as electrochemistry-based detection of post-PCR products.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Luo</LastName><ForeName>Xiaoteng</ForeName><Initials>X</Initials><Affiliation>Bioengineering Graduate Program, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.</Affiliation></Author><Author ValidYN="Y"><LastName>Lee</LastName><ForeName>Thomas Ming-Hung</ForeName><Initials>TM</Initials></Author><Author ValidYN="Y"><LastName>Hsing</LastName><ForeName>I-Ming</ForeName><Initials>IM</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType>Journal Article</PublicationType><PublicationType>Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2008</Year><Month>09</Month><Day>05</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Anal Chem</MedlineTA><NlmUniqueID>0370536</NlmUniqueID><ISSNLinking>0003-2700</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>DNA, Complementary</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>Ferrous Compounds</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>Peptide Nucleic Acids</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>Polyamines</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance>Tin Compounds</NameOfSubstance></Chemical><Chemical><RegistryNumber>30551-89-4</RegistryNumber><NameOfSubstance>polyallylamine</NameOfSubstance></Chemical><Chemical><RegistryNumber>71243-84-0</RegistryNumber><NameOfSubstance>indium tin oxide</NameOfSubstance></Chemical><Chemical><RegistryNumber>U96PKG90JQ</RegistryNumber><NameOfSubstance>ferrocene</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName MajorTopicYN="N">Base Pair Mismatch</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">DNA, Complementary</DescriptorName><QualifierName MajorTopicYN="Y">analysis</QualifierName><QualifierName MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Electrochemical Techniques</DescriptorName><QualifierName MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Electrodes</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Ferrous Compounds</DescriptorName><QualifierName MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Peptide Nucleic Acids</DescriptorName><QualifierName MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Polyamines</DescriptorName><QualifierName MajorTopicYN="N">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Polymerase Chain Reaction</DescriptorName><QualifierName MajorTopicYN="N">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Substrate Specificity</DescriptorName></MeshHeading><MeshHeading><DescriptorName MajorTopicYN="N">Tin Compounds</DescriptorName><QualifierName MajorTopicYN="N">chemistry</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="aheadofprint"><Year>2008</Year><Month>9</Month><Day>05</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2008</Year><Month>9</Month><Day>6</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2008</Year><Month>12</Month><Day>17</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2008</Year><Month>9</Month><Day>6</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="doi">10.1021/ac8010236</ArticleId><ArticleId IdType="pubmed">18771276</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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