Volume 149, 2011

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

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.

Article information

Article type
Paper
Submitted
18 May 2010
Accepted
22 Jul 2010
First published
11 Oct 2010

Faraday Discuss., 2011,149, 11-22

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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