Amperometric DNA sensor using the pyrroquinoline quinone glucose dehydrogenase-avidin conjugate

Biosens Bioelectron. 2002 Dec;17(11-12):1075-80. doi: 10.1016/s0956-5663(02)00101-x.

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

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Avidin / chemistry
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Biosensing Techniques / instrumentation*
  • Biosensing Techniques / methods
  • Biotin / chemistry
  • Calibration
  • Carbon
  • DNA / analysis
  • DNA / chemistry
  • DNA Probes / analysis
  • DNA, Bacterial / analysis*
  • DNA, Bacterial / chemistry
  • Electrochemistry / instrumentation*
  • Electrochemistry / methods
  • Electrodes
  • Enzymes, Immobilized / chemistry
  • Equipment Design
  • Glucose / analysis
  • Glucose / chemistry
  • Glucose Dehydrogenases / chemistry*
  • Materials Testing / methods
  • Reference Values
  • Reproducibility of Results
  • Salmonella / genetics
  • Salmonella / metabolism
  • Sensitivity and Specificity

Substances

  • Bacterial Proteins
  • DNA Probes
  • DNA, Bacterial
  • Enzymes, Immobilized
  • Avidin
  • invA protein, Bacteria
  • Biotin
  • Carbon
  • DNA
  • Glucose Dehydrogenases
  • glucose dehydrogenase (pyrroloquinoline-quinone)
  • Glucose