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Amplified Detection of DNA Hybridization Using Post-Labelling With a Biotin-Modified Intercalator

Amplified Detection of DNA Hybridization Using Post-Labelling With a Biotin-Modified Intercalator

Magdalena Gebala, Gerhard Hartwich, Wolfgang Schuhmann

Faraday Discuss. 2011;149:11-22; discussion 63-77.

PMID: 21413171

Abstract:

A 32-electrode microelectrode array modified with a self-assembled monolayer of a thiolated DNA capture strand and 11-mercapto-l1-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 epost-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 via oxidation ofp-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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42411638	Avidin-Alkaline Phosphatase			Price

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