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

2023-05-26Journal article Restricted access
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dc.contributor.authorMøller, Peter
dc.contributor.authorAzqueta, Amaya
dc.contributor.authorSanz-Serrano, Julen
dc.contributor.authorBakuradze, Tamara
dc.contributor.authorRichling, Elke
dc.contributor.authorEyluel Bankoglu, Ezgi
dc.contributor.authorStopper, Helga
dc.contributor.authorClaudino Bastos, Victoria
dc.contributor.authorLangie, Sabine A.S.
dc.contributor.authorJensen, Annie
dc.contributor.authorScavone, Francesca
dc.contributor.authorGiovannelli, Lisa
dc.contributor.authorWojewódzka, Maria
dc.contributor.authorKruszewski, Marcin
dc.contributor.authorValdiglesias, Vanessa
dc.contributor.authorLaffon, Blanca
dc.contributor.authorCosta, Carla
dc.contributor.authorCosta, Solange
dc.contributor.authorTeixeira, João Paulo
dc.contributor.authorMarino, Mirko
dc.contributor.authorDel Bo, Cristian
dc.contributor.authorRiso, Patrizia
dc.contributor.authorZheng, Congying
dc.contributor.authorShaposhnikov, Sergey
dc.contributor.authorCollins, Andrew
dc.date.accessioned2024-02-08T16:49:54Z
dc.date.available2024-02-08T16:49:54Z
dc.date.issued2023-05-26
dc.description.abstractMeasurement 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.pt_PT
dc.description.sponsorshipCertain authors have acknowledged funding as follows: statutory funding for INTC and IRH (Maria Wojewódzka and Marcin Kruszewski); Spanish Ministry of Science and Innovation MCIN/AEI/10.13039/501100011033 (Grants PID2020- 113788RB-I00 and PID2020-114908 GA-I00), and Xunta de Galicia (Grant ED431B 2022/16) (Vanessa Valdiglesias and Blanca Laffon); Spanish Ministry of Science and Innovation (Ref.: PID2020-115348RB-I00) (Amaya Azqueta); Università degli Studi di Firenze fund RICATEN21 and RICATEN22 (Lisa Giovannelli).
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMutagenesis. 2023 Oct 14;38(5):253-263. doi: 10.1093/mutage/gead015pt_PT
dc.identifier.doi10.1093/mutage/gead015pt_PT
dc.identifier.issn0267-8357
dc.identifier.urihttp://hdl.handle.net/10400.18/9062
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherOxford University Press/ UK Environmental Mutagen Societypt_PT
dc.relation.publisherversionhttps://academic.oup.com/mutage/article-abstract/38/5/253/7180292?redirectedFrom=fulltext&login=truept_PT
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/pt_PT
dc.subjectComet Assaypt_PT
dc.subjectDNA Damagept_PT
dc.subjectRing Trialpt_PT
dc.subjectValidationpt_PT
dc.subjectVisual Scoringpt_PT
dc.subjectToxicologiapt_PT
dc.titleVisual comet scoring revisited: a guide to scoring comet assay slides and obtaining reliable resultspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage263pt_PT
oaire.citation.issue5pt_PT
oaire.citation.startPage253pt_PT
oaire.citation.titleMutagenesispt_PT
oaire.citation.volume38pt_PT
rcaap.embargofctAcesso de acordo com política editorial da revista.pt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT

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