Peter Møller, Amaya Azqueta, Julen Sanz-Serrano, Tamara Bakuradze, Elke Richling, Ezgi Eyluel Bankoglu, Helga Stopper, Victoria Claudino Bastos, Sabine A S Langie, Annie Jensen, Francesca Scavone, Lisa Giovannelli, Maria Wojewódzka, Marcin Kruszewski, Vanessa Valdiglesias, Blanca Laffon, Carla Costa, Solange Costa, João Paulo Teixeira, Mirko Marino, Cristian Del Bo, Patrizia Riso, Congying Zheng, Sergey Shaposhnikov, Andrew Collins
{"title":"再次访问视觉彗星评分:彗星分析幻灯片评分和获得可靠结果的指南。","authors":"Peter Møller, Amaya Azqueta, Julen Sanz-Serrano, Tamara Bakuradze, Elke Richling, Ezgi Eyluel Bankoglu, Helga Stopper, Victoria Claudino Bastos, Sabine A S Langie, Annie Jensen, Francesca Scavone, Lisa Giovannelli, Maria Wojewódzka, Marcin Kruszewski, Vanessa Valdiglesias, Blanca Laffon, Carla Costa, Solange Costa, João Paulo Teixeira, Mirko Marino, Cristian Del Bo, Patrizia Riso, Congying Zheng, Sergey Shaposhnikov, Andrew Collins","doi":"10.1093/mutage/gead015","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"253-263"},"PeriodicalIF":2.5000,"publicationDate":"2023-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"3","resultStr":"{\"title\":\"Visual comet scoring revisited: a guide to scoring comet assay slides and obtaining reliable results.\",\"authors\":\"Peter Møller, Amaya Azqueta, Julen Sanz-Serrano, Tamara Bakuradze, Elke Richling, Ezgi Eyluel Bankoglu, Helga Stopper, Victoria Claudino Bastos, Sabine A S Langie, Annie Jensen, Francesca Scavone, Lisa Giovannelli, Maria Wojewódzka, Marcin Kruszewski, Vanessa Valdiglesias, Blanca Laffon, Carla Costa, Solange Costa, João Paulo Teixeira, Mirko Marino, Cristian Del Bo, Patrizia Riso, Congying Zheng, Sergey Shaposhnikov, Andrew Collins\",\"doi\":\"10.1093/mutage/gead015\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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. 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Visual comet scoring revisited: a guide to scoring comet assay slides and obtaining reliable results.
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.
期刊介绍:
Mutagenesis is an international multi-disciplinary journal designed to bring together research aimed at the identification, characterization and elucidation of the mechanisms of action of physical, chemical and biological agents capable of producing genetic change in living organisms and the study of the consequences of such changes.