Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial
Autor(a) principal: | |
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Data de Publicação: | 2010 |
Outros Autores: | , , , , , , , , , , , , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.18/100 |
Resumo: | The comet assay has become a popular method for the assessment of DNA damage in biomonitoring studies and genetic toxicology. However, few studies have addressed the issue of the noted inter-laboratory variability of DNA damage measured by the comet assay. In this study, 12 laboratories analysed the level of DNA damage in monocyte-derived THP-1 cells by either visual classification or computer-aided image analysis of pre-made slides, coded cryopreserved samples of cells and reference standard cells (calibration curve samples). The reference standard samples were irradiated with ionizing radiation (0–10 Gy) and used to construct a calibration curve to calculate the number of lesions per 106 base pair. All laboratories detected dose–response relationships in the coded samples irradiated with ionizing radiation (1.5–7 Gy), but there were overt differences in the level of DNA damage reported by the different laboratories as evidenced by an inter-laboratory coefficient of variation (CV) of 47%. Adjustment of the primary comet assay end points by a calibration curve prepared in each laboratory reduced the CV to 28%, a statistically significant reduction (P < 0.05, Levene's test). A large fraction of the inter-laboratory variation originated from differences in image analysis, whereas the intra-laboratory variation was considerably smaller than the variation between laboratories. In summary, adjustment of primary comet assay results by reference standards reduces inter-laboratory variation in the level of DNA damage measured by the alkaline version of the comet assay. |
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Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trialSaúde ambientalAr e Saúde OcupacionalToxicologiaThe comet assay has become a popular method for the assessment of DNA damage in biomonitoring studies and genetic toxicology. However, few studies have addressed the issue of the noted inter-laboratory variability of DNA damage measured by the comet assay. In this study, 12 laboratories analysed the level of DNA damage in monocyte-derived THP-1 cells by either visual classification or computer-aided image analysis of pre-made slides, coded cryopreserved samples of cells and reference standard cells (calibration curve samples). The reference standard samples were irradiated with ionizing radiation (0–10 Gy) and used to construct a calibration curve to calculate the number of lesions per 106 base pair. All laboratories detected dose–response relationships in the coded samples irradiated with ionizing radiation (1.5–7 Gy), but there were overt differences in the level of DNA damage reported by the different laboratories as evidenced by an inter-laboratory coefficient of variation (CV) of 47%. Adjustment of the primary comet assay end points by a calibration curve prepared in each laboratory reduced the CV to 28%, a statistically significant reduction (P < 0.05, Levene's test). A large fraction of the inter-laboratory variation originated from differences in image analysis, whereas the intra-laboratory variation was considerably smaller than the variation between laboratories. In summary, adjustment of primary comet assay results by reference standards reduces inter-laboratory variation in the level of DNA damage measured by the alkaline version of the comet assay.Oxford University PressRepositório Científico do Instituto Nacional de SaúdeForchhammer, LykkeJohansson, ClaraLoft, SteffenMöller, LennartGodschalk, Roger, W. L.Langie, Sabine A. S.Jones, George D. D.Kwok, Rachel W. L.Collins, Andrew R.Azqueta, AmayaPhillips, David H.Sozeri, OsmanStepnik, MaciejPalus, JadwigaVogel, UllaWallin, HakanRoutledge, Michael N.Handforth, CatherineAllione, AlessandraMatullo, GiuseppeTeixeira, João PauloCosta, SolangeRiso, PatriziaPorrini, MarisaMøller, Peter2011-06-01T16:39:14Z2010-03-022010-03-02T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/100engMutagenesis. 2010 Mar;25(2):113-23. Epub 2009 Nov 120267-8357info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-20T15:38:00Zoai:repositorio.insa.pt:10400.18/100Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:35:18.040111Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial |
title |
Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial |
spellingShingle |
Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial Forchhammer, Lykke Saúde ambiental Ar e Saúde Ocupacional Toxicologia |
title_short |
Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial |
title_full |
Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial |
title_fullStr |
Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial |
title_full_unstemmed |
Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial |
title_sort |
Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial |
author |
Forchhammer, Lykke |
author_facet |
Forchhammer, Lykke Johansson, Clara Loft, Steffen Möller, Lennart Godschalk, Roger, W. L. Langie, Sabine A. S. Jones, George D. D. Kwok, Rachel W. L. Collins, Andrew R. Azqueta, Amaya Phillips, David H. Sozeri, Osman Stepnik, Maciej Palus, Jadwiga Vogel, Ulla Wallin, Hakan Routledge, Michael N. Handforth, Catherine Allione, Alessandra Matullo, Giuseppe Teixeira, João Paulo Costa, Solange Riso, Patrizia Porrini, Marisa Møller, Peter |
author_role |
author |
author2 |
Johansson, Clara Loft, Steffen Möller, Lennart Godschalk, Roger, W. L. Langie, Sabine A. S. Jones, George D. D. Kwok, Rachel W. L. Collins, Andrew R. Azqueta, Amaya Phillips, David H. Sozeri, Osman Stepnik, Maciej Palus, Jadwiga Vogel, Ulla Wallin, Hakan Routledge, Michael N. Handforth, Catherine Allione, Alessandra Matullo, Giuseppe Teixeira, João Paulo Costa, Solange Riso, Patrizia Porrini, Marisa Møller, Peter |
author2_role |
author author author author author author author author author author author author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório Científico do Instituto Nacional de Saúde |
dc.contributor.author.fl_str_mv |
Forchhammer, Lykke Johansson, Clara Loft, Steffen Möller, Lennart Godschalk, Roger, W. L. Langie, Sabine A. S. Jones, George D. D. Kwok, Rachel W. L. Collins, Andrew R. Azqueta, Amaya Phillips, David H. Sozeri, Osman Stepnik, Maciej Palus, Jadwiga Vogel, Ulla Wallin, Hakan Routledge, Michael N. Handforth, Catherine Allione, Alessandra Matullo, Giuseppe Teixeira, João Paulo Costa, Solange Riso, Patrizia Porrini, Marisa Møller, Peter |
dc.subject.por.fl_str_mv |
Saúde ambiental Ar e Saúde Ocupacional Toxicologia |
topic |
Saúde ambiental Ar e Saúde Ocupacional Toxicologia |
description |
The comet assay has become a popular method for the assessment of DNA damage in biomonitoring studies and genetic toxicology. However, few studies have addressed the issue of the noted inter-laboratory variability of DNA damage measured by the comet assay. In this study, 12 laboratories analysed the level of DNA damage in monocyte-derived THP-1 cells by either visual classification or computer-aided image analysis of pre-made slides, coded cryopreserved samples of cells and reference standard cells (calibration curve samples). The reference standard samples were irradiated with ionizing radiation (0–10 Gy) and used to construct a calibration curve to calculate the number of lesions per 106 base pair. All laboratories detected dose–response relationships in the coded samples irradiated with ionizing radiation (1.5–7 Gy), but there were overt differences in the level of DNA damage reported by the different laboratories as evidenced by an inter-laboratory coefficient of variation (CV) of 47%. Adjustment of the primary comet assay end points by a calibration curve prepared in each laboratory reduced the CV to 28%, a statistically significant reduction (P < 0.05, Levene's test). A large fraction of the inter-laboratory variation originated from differences in image analysis, whereas the intra-laboratory variation was considerably smaller than the variation between laboratories. In summary, adjustment of primary comet assay results by reference standards reduces inter-laboratory variation in the level of DNA damage measured by the alkaline version of the comet assay. |
publishDate |
2010 |
dc.date.none.fl_str_mv |
2010-03-02 2010-03-02T00:00:00Z 2011-06-01T16:39:14Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.18/100 |
url |
http://hdl.handle.net/10400.18/100 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Mutagenesis. 2010 Mar;25(2):113-23. Epub 2009 Nov 12 0267-8357 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Oxford University Press |
publisher.none.fl_str_mv |
Oxford University Press |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799132078752661504 |