Inter-laboratory variation in DNA damage using a standard comet assay protocol.

2.50
Hdl Handle:
http://hdl.handle.net/10146/237014
Title:
Inter-laboratory variation in DNA damage using a standard comet assay protocol.
Authors:
Forchhammer, Lykke; Ersson, Clara; Loft, Steffen; Moller, Lennart; Godschalk, Roger W. L.; van Schooten, Frederik J.; Jones, George D. D.; Higgins, Jennifer A.; Cooke, Marcus; Mistry, Vilas; Karbaschi, Mahsa; Collins, Andrew R.; Azqueta, Amaya; Phillips, David H.; Sozeri, Osman; Routledge, Michael N.; Nelson-Smith, Kirsty; Riso, Patrizia; Porrini, Marisa; Matullo, Giuseppe; Allione, Alessandra; Stepnik, Maciej; Komorowska, Magdalena; Teixeira, Joao Paulo; Costa, Solange; Corcuera, Laura-Ana; Lopez de Cerain, Adela; Laffon, Blanca; Valdiglesias, Vanessa; Moller, Peter
Abstract:
There are substantial inter-laboratory variations in the levels of DNA damage measured by the comet assay. The aim of this study was to investigate whether adherence to a standard comet assay protocol would reduce inter-laboratory variation in reported values of DNA damage. Fourteen laboratories determined the baseline level of DNA strand breaks (SBs)/alkaline labile sites and formamidopyrimidine DNA glycosylase (FPG)-sensitive sites in coded samples of mononuclear blood cells (MNBCs) from healthy volunteers. There were technical problems in seven laboratories in adopting the standard protocol, which were not related to the level of experience. Therefore, the inter-laboratory variation in DNA damage was only analysed using the results from laboratories that had obtained complete data with the standard comet assay protocol. This analysis showed that the differences between reported levels of DNA SBs/alkaline labile sites in MNBCs were not reduced by applying the standard assay protocol as compared with the laboratory's own protocol. There was large inter-laboratory variation in FPG-sensitive sites by the laboratory-specific protocol and the variation was reduced when the samples were analysed by the standard protocol. The SBs and FPG-sensitive sites were measured in the same experiment, indicating that the large spread in the latter lesions was the main reason for the reduced inter-laboratory variation. However, it remains worrying that half of the participating laboratories obtained poor results using the standard procedure. This study indicates that future comet assay validation trials should take steps to evaluate the implementation of standard procedures in participating laboratories.
Citation:
Mutagenesis 2012, 27 (6):665-672
Journal:
Mutagenesis
Issue Date:
27-Jul-2012
URI:
http://hdl.handle.net/10146/237014
DOI:
10.1093/mutage/ges032
PubMed ID:
22844078
Additional Links:
http://mutage.oxfordjournals.org/content/early/2012/07/21/mutage.ges032.long
Type:
Article; Preprint
Language:
en
ISSN:
1464-3804
Sponsors:
This work was supported by ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6th Framework Program, Priority 5: “Food Quality and Safety” (contract no. 513943).
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorForchhammer, Lykkeen_GB
dc.contributor.authorErsson, Claraen_GB
dc.contributor.authorLoft, Steffenen_GB
dc.contributor.authorMoller, Lennarten_GB
dc.contributor.authorGodschalk, Roger W. L.en_GB
dc.contributor.authorvan Schooten, Frederik J.en_GB
dc.contributor.authorJones, George D. D.en_GB
dc.contributor.authorHiggins, Jennifer A.en_GB
dc.contributor.authorCooke, Marcusen_GB
dc.contributor.authorMistry, Vilasen_GB
dc.contributor.authorKarbaschi, Mahsaen_GB
dc.contributor.authorCollins, Andrew R.en_GB
dc.contributor.authorAzqueta, Amayaen_GB
dc.contributor.authorPhillips, David H.en_GB
dc.contributor.authorSozeri, Osmanen_GB
dc.contributor.authorRoutledge, Michael N.en_GB
dc.contributor.authorNelson-Smith, Kirstyen_GB
dc.contributor.authorRiso, Patriziaen_GB
dc.contributor.authorPorrini, Marisaen_GB
dc.contributor.authorMatullo, Giuseppeen_GB
dc.contributor.authorAllione, Alessandraen_GB
dc.contributor.authorStepnik, Maciejen_GB
dc.contributor.authorKomorowska, Magdalenaen_GB
dc.contributor.authorTeixeira, Joao Pauloen_GB
dc.contributor.authorCosta, Solangeen_GB
dc.contributor.authorCorcuera, Laura-Anaen_GB
dc.contributor.authorLopez de Cerain, Adelaen_GB
dc.contributor.authorLaffon, Blancaen_GB
dc.contributor.authorValdiglesias, Vanessaen_GB
dc.contributor.authorMoller, Peteren_GB
dc.date.accessioned2012-08-02T10:24:31Z-
dc.date.available2012-08-02T10:24:31Z-
dc.date.issued2012-07-27-
dc.identifier.citationMutagenesis 2012, 27 (6):665-672en_GB
dc.identifier.issn1464-3804-
dc.identifier.pmid22844078-
dc.identifier.doi10.1093/mutage/ges032-
dc.identifier.urihttp://hdl.handle.net/10146/237014-
dc.description.abstractThere are substantial inter-laboratory variations in the levels of DNA damage measured by the comet assay. The aim of this study was to investigate whether adherence to a standard comet assay protocol would reduce inter-laboratory variation in reported values of DNA damage. Fourteen laboratories determined the baseline level of DNA strand breaks (SBs)/alkaline labile sites and formamidopyrimidine DNA glycosylase (FPG)-sensitive sites in coded samples of mononuclear blood cells (MNBCs) from healthy volunteers. There were technical problems in seven laboratories in adopting the standard protocol, which were not related to the level of experience. Therefore, the inter-laboratory variation in DNA damage was only analysed using the results from laboratories that had obtained complete data with the standard comet assay protocol. This analysis showed that the differences between reported levels of DNA SBs/alkaline labile sites in MNBCs were not reduced by applying the standard assay protocol as compared with the laboratory's own protocol. There was large inter-laboratory variation in FPG-sensitive sites by the laboratory-specific protocol and the variation was reduced when the samples were analysed by the standard protocol. The SBs and FPG-sensitive sites were measured in the same experiment, indicating that the large spread in the latter lesions was the main reason for the reduced inter-laboratory variation. However, it remains worrying that half of the participating laboratories obtained poor results using the standard procedure. This study indicates that future comet assay validation trials should take steps to evaluate the implementation of standard procedures in participating laboratories.en_GB
dc.description.sponsorshipThis work was supported by ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6th Framework Program, Priority 5: “Food Quality and Safety” (contract no. 513943).en_GB
dc.languageENG-
dc.language.isoenen
dc.relation.urlhttp://mutage.oxfordjournals.org/content/early/2012/07/21/mutage.ges032.longen_GB
dc.rightsArchived with thanks to Mutagenesisen_GB
dc.subjectDNA damageen_GB
dc.subjectBiomarkersen_GB
dc.subjectComet assayen_GB
dc.subjectHumanen_GB
dc.subjectLaboratoriesen_GB
dc.subjectValidationen_GB
dc.subject.meshCalibration-
dc.subject.meshComet Assay-
dc.subject.meshDNA Damage-
dc.subject.meshDNA-Formamidopyrimidine Glycosylase-
dc.subject.meshEndpoint Determination-
dc.subject.meshHumans-
dc.subject.meshLaboratories-
dc.subject.meshLeukocytes, Mononuclear-
dc.subject.meshLinear Models-
dc.titleInter-laboratory variation in DNA damage using a standard comet assay protocol.en
dc.typeArticleen
dc.typePreprinten
dc.identifier.journalMutagenesisen_GB

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