Variation in the measurement of DNA damage by comet assay measured by the ECVAG{dagger} inter-laboratory validation trial
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Variation in the measurement of DNA damage by comet assay measured by the ECVAG{dagger} inter-laboratory validation trial. / 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, Håkan; Routledge, Michael N; Handforth, Catherine; Allione, Alessandra; Matullo, Giuseppe; Teixeira, João Paulo; Costa, Solange; Riso, Patrizia; Porrini, Marisa; Møller, Peter.
In: Mutagenesis, Vol. 25, No. 2, 2010, p. 113-23.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Variation in the measurement of DNA damage by comet assay measured by the ECVAG{dagger} inter-laboratory validation trial
AU - Forchhammer, Lykke
AU - Johansson, Clara
AU - Loft, Steffen
AU - Möller, Lennart
AU - Godschalk, Roger W L
AU - Langie, Sabine A S
AU - Jones, George D D
AU - Kwok, Rachel W L
AU - Collins, Andrew R
AU - Azqueta, Amaya
AU - Phillips, David H
AU - Sozeri, Osman
AU - Stepnik, Maciej
AU - Palus, Jadwiga
AU - Vogel, Ulla
AU - Wallin, Håkan
AU - Routledge, Michael N
AU - Handforth, Catherine
AU - Allione, Alessandra
AU - Matullo, Giuseppe
AU - Teixeira, João Paulo
AU - Costa, Solange
AU - Riso, Patrizia
AU - Porrini, Marisa
AU - Møller, Peter
PY - 2010
Y1 - 2010
N2 - 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 10(6) 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.
AB - 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 10(6) 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.
U2 - 10.1093/mutage/gep048
DO - 10.1093/mutage/gep048
M3 - Journal article
C2 - 19910383
VL - 25
SP - 113
EP - 123
JO - Mutagenesis
JF - Mutagenesis
SN - 0267-8357
IS - 2
ER -
ID: 18360127