Reliable Comet assay measurements for detecting DNA damage induced by ionising radiation and chemicals

Mutation Research
T S Kumaravel, Awadhesh N Jha

Abstract

The alkaline version of the single cell gel electrophoresis assay, popularly known as the Comet assay, is widely used to evaluate the genotoxic potential of chemicals and environmental contaminants, and for environmental monitoring purposes. In recent years, this assay has increasingly been recognized as a potentially valuable tool for regulatory studies. The assay commonly utilises commercially available software programmes to evaluate the extent of DNA damage at the single-cell level. These programmes provide a large number of measurement outcomes (i.e., tail length, %Tail DNA, various measures of tail moment, etc.) to evaluate the extent of DNA migration and DNA damage. At the moment, however, there is no general agreement with respect to the most relevant measurements or parameters to use. This study was carried out to establish which measurement(s) in the Comet assay are most significantly correlated with DNA damage, and should thus be adopted for routine use. Pooled peripheral blood samples from 3 healthy human individuals were irradiated with a range of doses of (137)Cs gamma-radiation (0, 1, 2, 4 and 8 Gy). Following irradiation, the Comet assay was performed according to a standard protocol, and different parameters we...Continue Reading

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