Pulsed amperometric detection of DNA with an ssDNA/polypyrrole-modified electrode.
Identifieur interne : 002243 ( PubMed/Checkpoint ); précédent : 002242; suivant : 002244Pulsed amperometric detection of DNA with an ssDNA/polypyrrole-modified electrode.
Auteurs : Almira Ramanaviciene [Lituanie] ; Arunas RamanaviciusSource :
- Analytical and bioanalytical chemistry [ 1618-2642 ] ; 2004.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- Animals, Biosensing Techniques (instrumentation), Biosensing Techniques (methods), Cattle, DNA (analysis), DNA, Single-Stranded (chemistry), DNA, Viral (analysis), Electrochemistry (instrumentation), Electrochemistry (methods), Electrodes, Leukemia Virus, Bovine (chemistry), Polymers (chemistry), Pyrroles (chemistry).
- MESH :
- chemical , analysis : DNA, DNA, Viral.
- chemical , chemistry : DNA, Single-Stranded, Polymers, Pyrroles.
- chemistry : Leukemia Virus, Bovine.
- instrumentation : Biosensing Techniques, Electrochemistry.
- methods : Biosensing Techniques, Electrochemistry.
- Animals, Cattle, Electrodes.
Abstract
Pulsed amperometric detection (PAD) of target DNA with platinum electrodes modified by single-stranded DNA (ssDNA) entrapped within polypyrrole (ssDNA/Ppy) is reported for the first time. Single-stranded DNA 20-mers complementary to the target DNA were used to construct the DNA biosensors. Polymerase chain reaction (PCR) amplified bovine leukaemia virus (BLV) provirus DNA was used as target DNA. Electrochemical impedance spectroscopic (EIS) investigation of ssDNA/Ppy before and after incubation in target DNA-containing sample revealed significant changes in terms of an imaginary (Z") vs. a real (Z') component. The PAD results were in good agreement with EIS investigations. The PAD method was selected, because it does not require such sophisticated equipment as it is used to perform EIS and the results obtained can be more easily estimated. Optimum conditions for performing PAD and evaluating an analytical signal were elaborated. No label-binding step was necessary for detection of target DNA in PCR-amplified amplicons and detection time was reduced by as much as 30-35 min. The changes of PAD signals were at least 6-7 times higher if ssDNA/Ppy-modified electrodes instead of blank Ppy-modified electrodes were incubated in the target DNA solutions. If ssDNA/Ppy modified electrodes were incubated in non-complementary (control) DNA solution changes in PAD signals were smaller than those detected after incubation in complementary (target) DNA-containing solution by a factor of at least 6-8.
DOI: 10.1007/s00216-004-2573-6
PubMed: 15042269
Affiliations:
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pubmed:15042269Le document en format XML
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<term>Techniques de biocapteur (instrumentation)</term>
<term>Virus de la leucémie bovine ()</term>
<term>Électrochimie ()</term>
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<front><div type="abstract" xml:lang="en">Pulsed amperometric detection (PAD) of target DNA with platinum electrodes modified by single-stranded DNA (ssDNA) entrapped within polypyrrole (ssDNA/Ppy) is reported for the first time. Single-stranded DNA 20-mers complementary to the target DNA were used to construct the DNA biosensors. Polymerase chain reaction (PCR) amplified bovine leukaemia virus (BLV) provirus DNA was used as target DNA. Electrochemical impedance spectroscopic (EIS) investigation of ssDNA/Ppy before and after incubation in target DNA-containing sample revealed significant changes in terms of an imaginary (Z") vs. a real (Z') component. The PAD results were in good agreement with EIS investigations. The PAD method was selected, because it does not require such sophisticated equipment as it is used to perform EIS and the results obtained can be more easily estimated. Optimum conditions for performing PAD and evaluating an analytical signal were elaborated. No label-binding step was necessary for detection of target DNA in PCR-amplified amplicons and detection time was reduced by as much as 30-35 min. The changes of PAD signals were at least 6-7 times higher if ssDNA/Ppy-modified electrodes instead of blank Ppy-modified electrodes were incubated in the target DNA solutions. If ssDNA/Ppy modified electrodes were incubated in non-complementary (control) DNA solution changes in PAD signals were smaller than those detected after incubation in complementary (target) DNA-containing solution by a factor of at least 6-8.</div>
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<Abstract><AbstractText>Pulsed amperometric detection (PAD) of target DNA with platinum electrodes modified by single-stranded DNA (ssDNA) entrapped within polypyrrole (ssDNA/Ppy) is reported for the first time. Single-stranded DNA 20-mers complementary to the target DNA were used to construct the DNA biosensors. Polymerase chain reaction (PCR) amplified bovine leukaemia virus (BLV) provirus DNA was used as target DNA. Electrochemical impedance spectroscopic (EIS) investigation of ssDNA/Ppy before and after incubation in target DNA-containing sample revealed significant changes in terms of an imaginary (Z") vs. a real (Z') component. The PAD results were in good agreement with EIS investigations. The PAD method was selected, because it does not require such sophisticated equipment as it is used to perform EIS and the results obtained can be more easily estimated. Optimum conditions for performing PAD and evaluating an analytical signal were elaborated. No label-binding step was necessary for detection of target DNA in PCR-amplified amplicons and detection time was reduced by as much as 30-35 min. The changes of PAD signals were at least 6-7 times higher if ssDNA/Ppy-modified electrodes instead of blank Ppy-modified electrodes were incubated in the target DNA solutions. If ssDNA/Ppy modified electrodes were incubated in non-complementary (control) DNA solution changes in PAD signals were smaller than those detected after incubation in complementary (target) DNA-containing solution by a factor of at least 6-8.</AbstractText>
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