Amperometric DNA sensor using the pyrroquinoline quinone glucose dehydrogenase-avidin conjugate.
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.
Ikebukuro
Kazunori
K
Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-machi, Koganei-shi, Tokyo 184-8588, Japan.
Kohiki
Yumiko
Y
Sode
Koji
K
D001360:Avidin / Q000737:chemistry; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D001710:Biotin / Q000737:chemistry; D002138:Calibration; D002244:Carbon; D004247:DNA / Q000032:analysis / Q000737:chemistry; D015342:DNA Probes / Q000032:analysis; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry; D004867:Equipment Design; D005947:Glucose / Q000032:analysis / Q000737:chemistry; D005948:Glucose Dehydrogenases / Q000737:chemistry*; D008422:Materials Testing / Q000379:methods; D012016:Reference Values; D015203:Reproducibility of Results; D012475:Salmonella / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity
D001426:Bacterial Proteins; D015342:DNA Probes; D004269:DNA, Bacterial; D004800:Enzymes, Immobilized; D001360:Avidin; C075518:invA protein, Bacteria; D001710:Biotin; D002244:Carbon; D004247:DNA; D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone); D005947:Glucose