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Sensors 2008, 8(1), 193-210; doi:10.3390/s8010193
Article

Enzyme-Linked Electrochemical Detection of PCR-Amplified Nucleotide Sequences Using Disposable Screen-Printed Sensors. Applications in Gene Expression Monitoring

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Received: 28 December 2007; Accepted: 7 January 2008 / Published: 21 January 2008
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Abstract: Electrochemical enzyme-linked techniques for sequence-specific DNA sensingare presented. These techniques are based on attachment of streptavidin-alkalinephosphatase conjugate to biotin tags tethered to DNA immobilized at the surface ofdisposable screen-printed carbon electrodes (SPCE), followed by production andelectrochemical determination of an electroactive indicator, 1-naphthol. Via hybridizationof SPCE surface-confined target DNAs with end-biotinylated probes, highly specificdiscrimination between complementary and non-complementary nucleotide sequences wasachieved. The enzyme-linked DNA hybridization assay has been successfully applied inanalysis of PCR-amplified real genomic DNA sequences, as well as in monitoring of planttissue-specific gene expression. In addition, we present an alternative approach involvingsequence-specific incorporation of biotin-labeled nucleotides into DNA by primerextension. Introduction of multiple biotin tags per probe primer resulted in considerableenhancement of the signal intensity and improvement of the specificity of detection.
Keywords: electrochemical detection; enzyme-linked assay; DNA hybridization; primer extension; PCR; gene expression electrochemical detection; enzyme-linked assay; DNA hybridization; primer extension; PCR; gene expression
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Horaková-Brazdilova, P.; Fojtova, M.; Vytras, K.; Fojta, M. Enzyme-Linked Electrochemical Detection of PCR-Amplified Nucleotide Sequences Using Disposable Screen-Printed Sensors. Applications in Gene Expression Monitoring. Sensors 2008, 8, 193-210.

AMA Style

Horaková-Brazdilova P, Fojtova M, Vytras K, Fojta M. Enzyme-Linked Electrochemical Detection of PCR-Amplified Nucleotide Sequences Using Disposable Screen-Printed Sensors. Applications in Gene Expression Monitoring. Sensors. 2008; 8(1):193-210.

Chicago/Turabian Style

Horaková-Brazdilova, Petra; Fojtova, Miloslava; Vytras, Karel; Fojta, Miroslav. 2008. "Enzyme-Linked Electrochemical Detection of PCR-Amplified Nucleotide Sequences Using Disposable Screen-Printed Sensors. Applications in Gene Expression Monitoring." Sensors 8, no. 1: 193-210.



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