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 H(2)O(2) 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 approximately 1 x 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.
Pavlov
Valeri
V
The Institute of Chemistry, The Farkas Center for Light-Induced Processes, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Xiao
Yi
Y
Gill
Ron
R
Dishon
Arnon
A
Kotler
Moshe
M
Willner
Itamar
I
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D021881:DNA, Catalytic / Q000737:chemistry*; D006046:Gold / Q000494:pharmacology; D006367:HeLa Cells; D006427:Hemin / Q000378:metabolism; D006801:Humans; D049449:Luminescence; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D019920:Photoaffinity Labels / Q000737:chemistry; D013499:Surface Properties; D019098:Telomerase / Q000032:analysis* / Q000378:metabolism
D021881:DNA, Catalytic; D019920:Photoaffinity Labels; D006427:Hemin; D006046:Gold; D004247:DNA; D019098:Telomerase