Comet assay in human biomonitoring studies: Reliability, validation, and applications

TLDR

The comet assay, a single‑cell gel electrophoresis technique that quantifies DNA strand breaks, is easily applied to human lymphocytes and is therefore well suited for biomonitoring studies. This study evaluates the reliability and reproducibility of the comet assay, from duplicate gel comparisons to assessing natural intra‑ and interindividual variability in DNA damage among healthy volunteers. The assay was modified to detect DNA base oxidation by incorporating a lesion‑specific endonuclease step, performed on duplicate gels, and applied to investigations of disease and occupational exposure in factory workers. Published in Environmental Molecular Mutagenesis, vol.

Abstract

The comet assay (single-cell gel electrophoresis), which measures DNA strand breaks at the level of single cells, is very easily applied to human lymphocytes, and therefore lends itself to human biomonitoring studies. For the examination of DNA base oxidation (a specific marker of oxidative damage), the assay is modified by including a stage at which the DNA is incubated with a suitable lesion-specific endonuclease. Here we report on the reliability and reproducibility of this approach, from the level of comparing results from duplicate gels prepared from the same sample of cells, up to an assessment of the natural intra- and interindividual variability in lymphocyte DNA damage measured in groups of normal, healthy human volunteers. We applied the assay in investigations of human disease and occupational exposure of factory workers. Environ. Mol. Mutagen. 30:139–146, 1997. © 1997 Wiley-Liss, Inc.