Visual comet scoring revisited: a guide to scoring comet assay slides and obtaining reliable results.

Autor:	Møller P; Department of Public Health, Section of Environmental Health, University of Copenhagen, Øster Farimagsgade 5A, DK-1014 Copenhagen K, Denmark., Azqueta A; Department of Pharmacology and Toxicology, School of Pharmacy and Nutrition. University of Navarra, C/Irunlarrea 1, 31009 Pamplona, Spain., Sanz-Serrano J; Department of Pharmacology and Toxicology, School of Pharmacy and Nutrition. University of Navarra, C/Irunlarrea 1, 31009 Pamplona, Spain., Bakuradze T; Food Chemistry and Toxicology, Department of Chemistry, RPTU Kaiserslautern-Landau, Erwin-Schroedinger-Str. 52, D-67663 Kaiserslautern, Germany., Richling E; Food Chemistry and Toxicology, Department of Chemistry, RPTU Kaiserslautern-Landau, Erwin-Schroedinger-Str. 52, D-67663 Kaiserslautern, Germany., Eyluel Bankoglu E; Institute of Pharmacology and Toxicology, University of Würzburg, Versbacher Str. 9, 97078 Wuerzburg, Germany., Stopper H; Institute of Pharmacology and Toxicology, University of Würzburg, Versbacher Str. 9, 97078 Wuerzburg, Germany., Claudino Bastos V; Department of Pharmacology & Toxicology, School for Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University, Maastricht, The Netherlands., Langie SAS; Department of Pharmacology & Toxicology, School for Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University, Maastricht, The Netherlands., Jensen A; Department of Public Health, Section of Environmental Health, University of Copenhagen, Øster Farimagsgade 5A, DK-1014 Copenhagen K, Denmark., Scavone F; Department of Neuroscience, Psychology, Pharmacology and Child Health (NEUROFARBA), Section Pharmacology and Toxicology, University of Florence, Florence, Italy., Giovannelli L; Department of Neuroscience, Psychology, Pharmacology and Child Health (NEUROFARBA), Section Pharmacology and Toxicology, University of Florence, Florence, Italy., Wojewódzka M; Centre for Radiobiology and Biological Dosimetry, Institute of Nuclear Chemistry and Technology, Dorodna 16, 01-310 Warsaw, Poland., Kruszewski M; Centre for Radiobiology and Biological Dosimetry, Institute of Nuclear Chemistry and Technology, Dorodna 16, 01-310 Warsaw, Poland.; Department of Molecular Biology and Translational Research, Institute of Rural Health, Jaczewskiego 2, 20-090 Lublin, Poland., Valdiglesias V; Universidade da Coruña, Grupo NanoToxGen, Centro Interdisciplinar de Química e Bioloxía - CICA, Departamento de Biología, A Coruña, Spain.; Instituto de Investigación Biomédica de A Coruña (INIBIC), A Coruña, Spain., Laffon B; Instituto de Investigación Biomédica de A Coruña (INIBIC), A Coruña, Spain.; Universidade da Coruña, Grupo DICOMOSA, Centro Interdisciplinar de Química e Bioloxía - CICA, Departamento de Psicología, A Coruña, Spain., Costa C; Environmental Health Department, National Institute of Health, Porto, Portugal.; EPIUnit - Instituto de Saúde Pública, Universidade do Porto, Porto, Portugal.; Laboratory for Integrative and Translational Research in Population Health (ITR), Porto, Portugal., Costa S; Environmental Health Department, National Institute of Health, Porto, Portugal.; EPIUnit - Instituto de Saúde Pública, Universidade do Porto, Porto, Portugal.; Laboratory for Integrative and Translational Research in Population Health (ITR), Porto, Portugal., Teixeira JP; Environmental Health Department, National Institute of Health, Porto, Portugal.; EPIUnit - Instituto de Saúde Pública, Universidade do Porto, Porto, Portugal.; Laboratory for Integrative and Translational Research in Population Health (ITR), Porto, Portugal., Marino M; Department of Food, Environmental and Nutritional Sciences (DeFENS), Università degli Studi di Milano, 20133 Milan, Italy., Del Bo C; Department of Food, Environmental and Nutritional Sciences (DeFENS), Università degli Studi di Milano, 20133 Milan, Italy., Riso P; Department of Food, Environmental and Nutritional Sciences (DeFENS), Università degli Studi di Milano, 20133 Milan, Italy., Zheng C; Department of Pharmacology & Toxicology, School for Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University, Maastricht, The Netherlands.; NorGenotech AS, Oslo, Norway., Shaposhnikov S; NorGenotech AS, Oslo, Norway., Collins A; NorGenotech AS, Oslo, Norway.; Department of Nutrition, University of Oslo, Oslo, Norway.
Jazyk:	angličtina
Zdroj:	Mutagenesis [Mutagenesis] 2023 Oct 14; Vol. 38 (5), pp. 253-263.
DOI:	10.1093/mutage/gead015
Abstrakt:	Measurement of DNA migration in the comet assay can be done by image analysis or visual scoring. The latter accounts for 20%-25% of the published comet assay results. Here we assess the intra- and inter-investigator variability in visual scoring of comets. We include three training sets of comet images, which can be used as reference for researchers who wish to use visual scoring of comets. Investigators in 11 different laboratories scored the comet images using a five-class scoring system. There is inter-investigator variation in the three training sets of comets (i.e. coefficient of variation (CV) = 9.7%, 19.8%, and 15.2% in training sets I-III, respectively). However, there is also a positive correlation of inter-investigator scoring in the three training sets (r = 0.60). Overall, 36% of the variation is attributed to inter-investigator variation and 64% stems from intra-investigator variation in scoring between comets (i.e. the comets in training sets I-III look slightly different and this gives rise to heterogeneity in scoring). Intra-investigator variation in scoring was also assessed by repeated analysis of the training sets by the same investigator. There was larger variation when the training sets were scored over a period of six months (CV = 5.9%-9.6%) as compared to 1 week (CV = 1.3%-6.1%). A subsequent study revealed a high inter-investigator variation when premade slides, prepared in a central laboratory, were stained and scored by investigators in different laboratories (CV = 105% and 18%-20% in premade slides with comets from unexposed and hydrogen peroxide-exposed cells, respectively). The results indicate that further standardization of visual scoring is desirable. Nevertheless, the analysis demonstrates that visual scoring is a reliable way of analysing DNA migration in comets. (© The Author(s) 2023. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
Databáze:	MEDLINE
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