Comparison of comet assay parameters for estimation of genotoxicity by sum of ranking differences

Sunjog, K. and Kolarević, S. and Héberger, Károly and Gačić, Z. and Knežević-Vukčević, J. and Vuković-Gačić, B. and Lenhardt, M. (2013) Comparison of comet assay parameters for estimation of genotoxicity by sum of ranking differences. Analytical and Bioanalytical Chemistry, 405 (14). 4879-4885 . ISSN 1618-2642

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Abstract

Genotoxic potential of waters in six rivers and reservoirs from Serbia was monitored in different tissues of chub (Squalius cephalus L. 1758) with the alkaline comet assay. The comet assay, or a single cell gel electrophoresis, has a wide application as a simple and sensitive method for evaluating DNA damage in fish exposed to various xenobiotics in the aquatic environment. Three types of cells, erythrocytes, gills and liver cells were used for assessing of DNA damage. Images of randomly selected cells were analyzed with a fluorescence microscope Leica and image analysis by software (Comet Assay IV Image analysis system, PI, UK). Three parameters (tail length - l, tail intensity - i and Olive tail moment – m) were analyzed on 1750 nuclei per cell type. The procedure for sum of ranking differences (SRD) was implemented to compare different types of cells and different parameters for estimation of DNA damage. Regarding our nine different estimations of genotoxicity: tail length, intensity and moment in erythrocytes (rel, rei, rem), liver cells (rll, rli, rlm) and gill cells (rgl, rgi, rgm) SRD procedure has shown that the Olive tail moment and tail intensity are (almost) equally good parameters; the SRD value was lower for the tail moment and tail intensity than for tail length in case of all types of cells. The least reliable parameter was rel; close to the borderline case were rei, rll, and rgl (~5% probability of random ranking).

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