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Title: Variation in the measurement of DNA damage by comet assay measured by the ECVAG† inter-laboratory validation trial
Author: 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
Keywords: Saúde ambiental
Ar e Saúde Ocupacional
Issue Date: 2-Mar-2010
Publisher: Oxford University Press
Citation: Mutagenesis. 2010 Mar;25(2):113-23. Epub 2009 Nov 12
Abstract: 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.
Peer review: yes
ISSN: 0267-8357
Publisher Version:
Appears in Collections:DSA - Artigos em revistas internacionais

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