Rapid, sequence-specific detection of unpurified PCR amplicons via a reusable, electrochemical sensor.
We report an electrochemical method for the sequence-specific detection of unpurified amplification products of the gyrB gene of Salmonella typhimurium. Using an asymmetric PCR and the electrochemical E-DNA detection scheme, single-stranded amplicons were produced from as few as 90 gene copies and, without subsequent purification, rapidly identified. The detection is specific; the sensor does not respond when challenged with control oligonucleotides based on the gyrB genes of either Escherichia coli or various Shigella species. In contrast to existing sequence-specific optical- and capillary electrophoresis-based detection methods, the E-DNA sensor is fully electronic and requires neither cumbersome, expensive optics nor high voltage power supplies. Given these advantages, E-DNA sensors appear well suited for implementation in portable PCR microdevices directed at, for example, the rapid detection of pathogens.
Lai
Rebecca Y
RY
Center for Polymers and Organic Solids, Department of Chemistry and Biochemistry, University of California, Santa Barbara, CA 93106, USA.
Lagally
Eric T
ET
Lee
Sang-Ho
SH
Soh
H T
HT
Plaxco
Kevin W
KW
Heeger
Alan J
AJ
D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D027081:DNA Gyrase / Q000235:genetics; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004563:Electrochemistry; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D021141:Nucleic Acid Amplification Techniques; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012486:Salmonella typhimurium / Q000235:genetics; D012763:Shigella flexneri / Q000235:genetics; D012764:Shigella sonnei / Q000235:genetics; D013045:Species Specificity
D015342:DNA Probes; D004269:DNA, Bacterial; D027081:DNA Gyrase