Abstract
A new amperometric DNA sensor was constructed using a pyrroquinoline quinone glucose dehydrogenase ((PQQ)GDH) conjugated with avidin. Our aim was to specifically detect the DNA sequence of the invA virulence gene from the pathogenic bacterium Salmonella. Probe DNA with a sequence complementary to that of a specific fragment of the invA gene was immobilized onto a carbon paste electrode. After hybridization with biotinylated target DNA, (PQQ)GDH-avidin conjugate was added and the resulting electric current was measured. The electric current is generated from glucose oxidization catalyzed by (PQQ)GDH via 1-methoxyphenazine methosulphate (m-PMS) electron mediator. The sensor response increased with the addition of glucose and in the presence of 6.3 mM glucose the response increased with increasing DNA in the range 5.0x10(-8)-1.0x10(-5) M.
Publication types
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Comparative Study
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Evaluation Study
MeSH terms
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Avidin / chemistry
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Bacterial Proteins / genetics*
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Bacterial Proteins / metabolism
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Biosensing Techniques / instrumentation*
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Biosensing Techniques / methods
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Biotin / chemistry
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Calibration
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Carbon
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DNA / analysis
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DNA / chemistry
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DNA Probes / analysis
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DNA, Bacterial / analysis*
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DNA, Bacterial / chemistry
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Electrochemistry / instrumentation*
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Electrochemistry / methods
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Electrodes
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Enzymes, Immobilized / chemistry
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Equipment Design
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Glucose / analysis
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Glucose / chemistry
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Glucose Dehydrogenases / chemistry*
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Materials Testing / methods
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Reference Values
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Reproducibility of Results
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Salmonella / genetics
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Salmonella / metabolism
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Sensitivity and Specificity
Substances
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Bacterial Proteins
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DNA Probes
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DNA, Bacterial
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Enzymes, Immobilized
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Avidin
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invA protein, Bacteria
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Biotin
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Carbon
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DNA
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Glucose Dehydrogenases
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glucose dehydrogenase (pyrroloquinoline-quinone)
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Glucose