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More specifically, the assay detects DNA migration from agarose gel-embedded nucleoids, which depends on assay conditions and the level of DNA damage. Certain steps in the comet assay procedure have substantial impact on the magnitude of DNA migration (e.g. electric potential and time of electrophoresis). Inter-laboratory variation in DNA migration levels occurs because there is no agreement on optimal assay conditions or suitable assay controls. The purpose of the hCOMET ring trial was to test potassium bromate (KBrO3) as a positive control for the formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay. To this end, participating laboratories used semi-standardized protocols for cell culture (i.e. cell culture, KBrO3 exposure, and cryopreservation of cells) and comet assay procedures, whereas the data acquisition was not standardized (i.e. staining of comets and image analysis). Segregation of the total variation into partial standard deviation (SD) in % Tail DNA units indicates the importance of cell culture procedures (SD = 10.9), comet assay procedures (SD = 12.3), staining (SD = 7.9) and image analysis (SD = 0.5) on the overall inter-laboratory variation of DNA migration (SD = 18.2). Future studies should assess sources of variation in each of these steps. On the positive side, the hCOMET ring trial demonstrates that KBrO3 is a robust positive control for the Fpg-modified comet assay. In conclusion, the hCOMET ring trial has demonstrated a high reproducibility of detecting genotoxic effects by the comet assay, but inter-laboratory variation of DNA migration levels is a concern.<\/jats:p>","DOI":"10.1093\/mutage\/gead014","type":"journal-article","created":{"date-parts":[[2023,5,25]],"date-time":"2023-05-25T19:35:35Z","timestamp":1685043335000},"page":"283-294","source":"Crossref","is-referenced-by-count":12,"title":["Inter-laboratory variation in measurement of DNA damage by the alkaline comet assay in the hCOMET ring trial"],"prefix":"10.1093","volume":"38","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2021-1249","authenticated-orcid":false,"given":"Peter","family":"M\u00f8ller","sequence":"first","affiliation":[{"name":"Department of Public Health, Section of Environmental Health, University of Copenhagen , \u00d8ster Farimagsgade 5A, DK-1014 Copenhagen K , Denmark"}]},{"given":"Amaya","family":"Azqueta","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, School of Pharmacy and Nutrition. University of Navarra , C\/Irunlarrea 1, 31009 Pamplona , Spain"}]},{"given":"Miguel","family":"Collia","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, School of Pharmacy and Nutrition. University of Navarra , C\/Irunlarrea 1, 31009 Pamplona , Spain"}]},{"given":"Tamara","family":"Bakuradze","sequence":"additional","affiliation":[{"name":"Food Chemistry and Toxicology, Department of Chemistry, RPTU Kaiserslautern-Landau, Erwin-Schroedinger-Str. 52 , D-67663 Kaiserslautern , Germany"}]},{"given":"Elke","family":"Richling","sequence":"additional","affiliation":[{"name":"Food Chemistry and Toxicology, Department of Chemistry, RPTU Kaiserslautern-Landau, Erwin-Schroedinger-Str. 52 , D-67663 Kaiserslautern , Germany"}]},{"given":"Ezgi Eyluel","family":"Bankoglu","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology and Toxicology, University of Wuerzburg , Versbacher Str. 9, 97078 Wuerzburg , Germany"}]},{"given":"Helga","family":"Stopper","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology and Toxicology, University of Wuerzburg , Versbacher Str. 9, 97078 Wuerzburg , Germany"}]},{"given":"Victoria Claudino","family":"Bastos","sequence":"additional","affiliation":[{"name":"Department of Pharmacology & Toxicology, School for Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University , Maastricht , The Netherlands"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3288-7331","authenticated-orcid":false,"given":"Sabine A S","family":"Langie","sequence":"additional","affiliation":[{"name":"Department of Pharmacology & Toxicology, School for Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University , Maastricht , The Netherlands"}]},{"given":"Annie","family":"Jensen","sequence":"additional","affiliation":[{"name":"Department of Public Health, Section of Environmental Health, University of Copenhagen , \u00d8ster Farimagsgade 5A, DK-1014 Copenhagen K , Denmark"}]},{"given":"Sara","family":"Ristori","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Psychology, Pharmacology and Child Health (NEUROFARBA), Section Pharmacology and Toxicology, University of Florence , Florence , Italy"}]},{"given":"Francesca","family":"Scavone","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Psychology, Pharmacology and Child Health (NEUROFARBA), Section Pharmacology and Toxicology, University of Florence , Florence , Italy"}]},{"given":"Lisa","family":"Giovannelli","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Psychology, Pharmacology and Child Health (NEUROFARBA), Section Pharmacology and Toxicology, University of Florence , Florence , Italy"}]},{"given":"Maria","family":"Wojew\u00f3dzka","sequence":"additional","affiliation":[{"name":"Centre for Radiobiology and Biological Dosimetry, Institute of Nuclear Chemistry and Technology , Dorodna 16, 01-310 Warsaw , Poland"}]},{"given":"Marcin","family":"Kruszewski","sequence":"additional","affiliation":[{"name":"Centre for Radiobiology and Biological Dosimetry, Institute of Nuclear Chemistry and Technology , Dorodna 16, 01-310 Warsaw , Poland"},{"name":"Department of Molecular Biology and Translational Research, Institute of Rural Health , Jaczewskiego 2, 20-090 Lublin , Poland"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5572-1089","authenticated-orcid":false,"given":"Vanessa","family":"Valdiglesias","sequence":"additional","affiliation":[{"name":"Universidade da Coru\u00f1a , Grupo NanoToxGen, Centro Interdisciplinar de Qu\u00edmica e Biolox\u00eda \u2013 CICA, Departamento de Biolog\u00eda, A Coru\u00f1a , Spain"},{"name":"Instituto de Investigaci\u00f3n Biom\u00e9dica de A Coru\u00f1a (INIBIC) , A Coru\u00f1a , Spain"}]},{"given":"Blanca","family":"Laffon","sequence":"additional","affiliation":[{"name":"Instituto de Investigaci\u00f3n Biom\u00e9dica de A Coru\u00f1a (INIBIC) , A Coru\u00f1a , Spain"},{"name":"Universidade da Coru\u00f1a, Grupo DICOMOSA , Centro Interdisciplinar de Qu\u00edmica e Biolox\u00eda \u2013 CICA, Departamento de Psicolog\u00eda, A Coru\u00f1a , Spain"}]},{"given":"Carla","family":"Costa","sequence":"additional","affiliation":[{"name":"Environmental Health Department, National Institute of Health , Porto , Portugal"},{"name":"EPIUnit - Instituto de Sa\u00fade P\u00fablica, Universidade do Porto , Porto , Portugal"},{"name":"Laboratory for Integrative and Translational Research in Population Health (ITR) , Porto , Portugal"}]},{"given":"Solange","family":"Costa","sequence":"additional","affiliation":[{"name":"Environmental Health Department, National Institute of Health , Porto , Portugal"},{"name":"EPIUnit - Instituto de Sa\u00fade P\u00fablica, Universidade do Porto , Porto , Portugal"},{"name":"Laboratory for Integrative and Translational Research in Population Health (ITR) , Porto , Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8693-5250","authenticated-orcid":false,"given":"Jo\u00e3o Paulo","family":"Teixeira","sequence":"additional","affiliation":[{"name":"Environmental Health Department, National Institute of Health , Porto , Portugal"},{"name":"EPIUnit - Instituto de Sa\u00fade P\u00fablica, Universidade do Porto , Porto , Portugal"},{"name":"Laboratory for Integrative and Translational Research in Population Health (ITR) , Porto , Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9913-4380","authenticated-orcid":false,"given":"Mirko","family":"Marino","sequence":"additional","affiliation":[{"name":"Department of Food, Environmental and Nutritional Sciences (DeFENS), Universita\u0300 degli Studi di Milano , 20133 Milan , Italy"}]},{"given":"Cristian","family":"Del Bo","sequence":"additional","affiliation":[{"name":"Department of Food, Environmental and Nutritional Sciences (DeFENS), Universita\u0300 degli Studi di Milano , 20133 Milan , Italy"}]},{"given":"Patrizia","family":"Riso","sequence":"additional","affiliation":[{"name":"Department of Food, Environmental and Nutritional Sciences (DeFENS), Universita\u0300 degli Studi di Milano , 20133 Milan , Italy"}]},{"given":"Congying","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Pharmacology & Toxicology, School for Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University , Maastricht , The Netherlands"},{"name":"Department of Nutrition, University of Oslo , Oslo , Norway"}]},{"given":"Sergey","family":"Shaposhnikov","sequence":"additional","affiliation":[{"name":"NorGenotech AS , Oslo , Norway"}]},{"given":"Andrew","family":"Collins","sequence":"additional","affiliation":[{"name":"Department of Nutrition, University of Oslo , Oslo , Norway"},{"name":"NorGenotech AS , Oslo , Norway"}]}],"member":"286","published-online":{"date-parts":[[2023,5,25]]},"reference":[{"key":"2023101415015444500_CIT0001","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.mrgentox.2019.04.007","article-title":"Technical recommendations to perform the alkaline standard and enzyme-modified comet assay in human biomonitoring studies","volume":"843","author":"Azqueta","year":"2019","journal-title":"Mutat Res Genet Toxicol Environ Mutagen"},{"key":"2023101415015444500_CIT0002","doi-asserted-by":"crossref","first-page":"2129","DOI":"10.1093\/carcin\/23.12.2129","article-title":"Comparative analysis of baseline 8-oxo-7,8-dihydroguanine in mammalian cell DNA, by different methods in different laboratories: an approach to consensus","volume":"23","author":"ESCODD","year":"2002","journal-title":"Carcinogenesis"},{"key":"2023101415015444500_CIT0003","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1016\/S0891-5849(03)00041-8","article-title":"Measurement of DNA oxidation in human cells by chromatographic and enzymic methods","volume":"34","author":"ESCODD","year":"2003","journal-title":"Free Radic Biol Med"},{"key":"2023101415015444500_CIT0004","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1096\/fj.04-1767fje","article-title":"Establishing the background level of base oxidation in human lymphocyte DNA: results of an inter-laboratory validation study","volume":"19","author":"Gedik","year":"2005","journal-title":"FASEB J"},{"key":"2023101415015444500_CIT0005","first-page":"25","article-title":"Sensitivity and variability of visual scoring in the comet assay. 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