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FIBER OPTIC BIOSENSOR FOR DNA DAMAGE
Rogers, K R., A. B. Apostol, S. J. Madsen, AND C. W. Spencer. FIBER OPTIC BIOSENSOR FOR DNA DAMAGE. ANALYTICA CHIMICA ACTA 444(1):51-60, (2001).
The overall objective of this task is to develop rapid, cost-effective and scientifically sound techniques for measuring chemically induced DNA damage. This method is expected to provide the Agency with rapid, sensitive, and simple techniques that can be used among a panel of methods to determine the genotoxic potential of polluted samples.
This paper describes a fiber optic biosensor for the rapid and sensitive detection of radiation-induced or chemically-induced oxidative DNA damage. The assay is based on the hybridization and temperature-induced dissociation (melting curves) of synthetic oligonucleotides. The hybridization pair consists of a biotin labeled 38-mer oligonucleotide immobilized to a streptavidin-coated optical fiber and a fluorescently-labeled near-complementary (2 base mismatch) oligonucleotide reporter sequence. The hybridization-based assay detected 50 nM labeled probe and could be run up to ten times on the same fiber. Melting profiles were sensitive to high energy radiation and to 3-morpholinosydnonimine (SIN-1)-generated reactive decomposition products. The dynamic range of the assay for ionizing radiation extends from 20cGy to 1000 cGy. Oxidative damage induced by SIN-1 was measured over a concentration range of 250 microM to 3 mM.
The US Environmental Protection Agency (EPA), through its Office of Research and Development (ORD), funded this research through a competitive internal grant (to K. R. Rogers). It has been subject to the EPA's peer and administrative review has been approved for publication. Mention of trade names or commercial products does not constitute endorsement or recommendation by EPA for use.