Issue 10, 2011

A DNAzyme based label-free detection system for miniaturized assays

Abstract

Sensitive detection assays are a prerequisite for the analysis of small amounts of samples derived from biological material. There is a great demand for highly sensitive and robust detection techniques to analyze biomolecules. The combination of catalytic active DNA (DNAzyme) with a peroxidase activity with rolling circle amplification (RCA) is a promising alternative to common detection systems. The rolling circle amplification leads to a product with tandemly linked copies of DNAzymes. The continuous signal generation of the amplified DNAzymes results in an increased sensitivity. The combination of two amplification reactions, namely RCA and DNAzymes, results in increased signal intensity by a factor of 106. With this approach the labeling of samples can be avoided. The advantage of the introduced assay is the usage of nucleic acids as biosensors for the detection of biomolecules. Coupling of the analyte molecule to the detection molecules allows the direct detection of the analyte molecule. The described label-free hotpot assay has a broad potential field of applications. The hotpot assay can be adapted to detect and analyze RNA, DNA and proteins down to femtomolar concentrations in a miniaturized platform with a total reaction solution of 50 nl. The applicability of the assay for diagnostics and research will be shown with a focus on high throughput systems using a nano-well platform.

Graphical abstract: A DNAzyme based label-free detection system for miniaturized assays

Article information

Article type
Paper
Submitted
05 Apr 2011
Accepted
04 Jul 2011
First published
27 Jul 2011

Mol. BioSyst., 2011,7, 2882-2889

A DNAzyme based label-free detection system for miniaturized assays

D. M. Köster, D. Haselbach, H. Lehrach and H. Seitz, Mol. BioSyst., 2011, 7, 2882 DOI: 10.1039/C1MB05132F

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