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Volume 149, 2011
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Amplified detection of DNA hybridization using post-labelling with a biotin-modified intercalator

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Abstract

A 32-electrode microelectrode array modified with a self-assembled monolayer of a thiolated DNA capture strand and 11-mercapto-1-undecanol was used for the detection of multi-resistant Staphylococcus aureus (MRSA) upon hybridization of the complementary target DNA. In the proposed assay strategy the obtained double-stranded DNA (dsDNA) is at first non-covalently labeled by intercalation of a proflavine derivative which is functionalized via a flexible spacer with biotin moieties. Subsequent to this post-labelling a avidin/alkaline phosphatase conjugate is bound to the biotin moieties thus introducing a reporter group at sites bearing dsDNA. Hybridization and hence the presence of MRSA DNA is detected viaoxidation of p-aminophenol enzymatically generated from p-aminophenylphosphate. The assay strategy was carefully evaluated using ferrocene-modified target strands. An increase in sensitivity of the proposed label-free DNA assays based on a careful design of the sensing interface and the implemented enzymatic amplification was achieved.

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Publication details

The article was received on 18 May 2010, accepted on 22 Jul 2010 and first published on 11 Oct 2010


Article type: Paper
DOI: 10.1039/C005365A
Faraday Discuss., 2011,149, 11-22

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    Amplified detection of DNA hybridization using post-labelling with a biotin-modified intercalator

    M. Gębala, G. Hartwich and W. Schuhmann, Faraday Discuss., 2011, 149, 11
    DOI: 10.1039/C005365A

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