Amplified Chemiluminescence Surface Detection of DNA and Telomerase Activity Using Catalytic Nucleic Acid Labels

A G-rich nucleic acid sequence binds hemin and yields a biocatalytic complex (DNAzyme) of peroxidase activity, namely, the biocatalyzed generation of chemiluminescence in the presence of H2O2 and luminol. The DNAzyme is used as a label for the amplified detection of DNA, or for the analysis of telom...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2004-04, Vol.76 (7), p.2152-2156
Main Authors: Pavlov, Valeri, Xiao, Yi, Gill, Ron, Dishon, Arnon, Kotler, Moshe, Willner, Itamar
Format: Article
Language:English
Subjects:
Analytical biochemistry: general aspects, technics, instrumentation
Analytical chemistry
Analytical, structural and metabolic biochemistry
Base Sequence
Biological and medical sciences
Catalysis
Chemical and thermal methods
Chemistry
Deoxyribonucleic acid
DNA
DNA - analysis
DNA - chemistry
DNA - metabolism
DNA, Catalytic - chemistry
Enzymes
Exact sciences and technology
Fundamental and applied biological sciences. Psychology
Gold - pharmacology
HeLa Cells
Hemin - metabolism
Humans
Labeling
Luminescence
Luminescent Measurements
Molecular Sequence Data
Photoaffinity Labels - chemistry
Surface Properties
Telomerase - analysis
Telomerase - metabolism
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Summary:A G-rich nucleic acid sequence binds hemin and yields a biocatalytic complex (DNAzyme) of peroxidase activity, namely, the biocatalyzed generation of chemiluminescence in the presence of H2O2 and luminol. The DNAzyme is used as a label for the amplified detection of DNA, or for the analysis of telomerase activity in cancer cells, using chemiluminescence as an output signal. In one configuration, the analyzed DNA is hybridized with a primer nucleic acid that is associated with a Au surface, and the DNAzyme label is hybridized with the surface-confined analyte DNA. The DNA is analyzed with a detection limit of ∼1 × 10-9 M. In the second system, telomerase from HeLa cancer cells induces telomerization of a primer associated with a Au surface and the complementary DNAzyme units are hybridized with the telomere to yield the chemiluminescence. The detection limit of the system corresponds to 1000 HeLa cells in the analyzed sample.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac035219l