An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay. / Johansson, Clara; Møller, Peter; Forchhammer, Lykke; Loft, Steffen; Godschalk, Roger W L; Langie, Sabine A S; Lumeij, Stijn; Jones, George D D; Kwok, Rachel W L; Azqueta, Amaya; Phillips, David H; Sozeri, Osman; Routledge, Michael N; Charlton, Alexander J; Riso, Patrizia; Porrini, Marisa; Allione, Alessandra; Matullo, Giuseppe; Palus, Jadwiga; Stepnik, Maciej; Collins, Andrew R; Möller, Lennart.

In: Mutagenesis, Vol. 25, No. 2, 2010, p. 125-32.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Johansson, C, Møller, P, Forchhammer, L, Loft, S, Godschalk, RWL, Langie, SAS, Lumeij, S, Jones, GDD, Kwok, RWL, Azqueta, A, Phillips, DH, Sozeri, O, Routledge, MN, Charlton, AJ, Riso, P, Porrini, M, Allione, A, Matullo, G, Palus, J, Stepnik, M, Collins, AR & Möller, L 2010, 'An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay', Mutagenesis, vol. 25, no. 2, pp. 125-32. https://doi.org/10.1093/mutage/gep055

APA

Johansson, C., Møller, P., Forchhammer, L., Loft, S., Godschalk, R. W. L., Langie, S. A. S., ... Möller, L. (2010). An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay. Mutagenesis, 25(2), 125-32. https://doi.org/10.1093/mutage/gep055

Vancouver

Johansson C, Møller P, Forchhammer L, Loft S, Godschalk RWL, Langie SAS et al. An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay. Mutagenesis. 2010;25(2):125-32. https://doi.org/10.1093/mutage/gep055

Author

Johansson, Clara ; Møller, Peter ; Forchhammer, Lykke ; Loft, Steffen ; Godschalk, Roger W L ; Langie, Sabine A S ; Lumeij, Stijn ; Jones, George D D ; Kwok, Rachel W L ; Azqueta, Amaya ; Phillips, David H ; Sozeri, Osman ; Routledge, Michael N ; Charlton, Alexander J ; Riso, Patrizia ; Porrini, Marisa ; Allione, Alessandra ; Matullo, Giuseppe ; Palus, Jadwiga ; Stepnik, Maciej ; Collins, Andrew R ; Möller, Lennart. / An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay. In: Mutagenesis. 2010 ; Vol. 25, No. 2. pp. 125-32.

Bibtex

@article{89f90d60269411df8ed1000ea68e967b,
title = "An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay",
abstract = "The increasing use of single cell gel electrophoresis (the comet assay) highlights its popularity as a method for detecting DNA damage, including the use of enzymes for assessment of oxidatively damaged DNA. However, comparison of DNA damage levels between laboratories can be difficult due to differences in assay protocols (e.g. lysis conditions, enzyme treatment, the duration of the alkaline treatment and electrophoresis) and in the end points used for reporting results (e.g. {\%}DNA in tail, arbitrary units, tail moment and tail length). One way to facilitate comparisons is to convert primary comet assay end points to number of lesions/10(6) bp by calibration with ionizing radiation. The aim of this study was to investigate the inter-laboratory variation in assessment of oxidatively damaged DNA by the comet assay in terms of oxidized purines converted to strand breaks with formamidopyrimidine DNA glycosylase (FPG). Coded samples with DNA oxidation damage induced by treatment with different concentrations of photosensitizer (Ro 19-8022) plus light and calibration samples irradiated with ionizing radiation were distributed to the 10 participating laboratories to measure DNA damage using their own comet assay protocols. Nine of 10 laboratories reported the same ranking of the level of damage in the coded samples. The variation in assessment of oxidatively damaged DNA was largely due to differences in protocols. After conversion of the data to lesions/10(6) bp using laboratory-specific calibration curves, the variation between the laboratories was reduced. The contribution of the concentration of photosensitizer to the variation in net FPG-sensitive sites increased from 49 to 73{\%}, whereas the inter-laboratory variation decreased. The participating laboratories were successful in finding a dose-response of oxidatively damaged DNA in coded samples, but there remains a need to standardize the protocols to enable direct comparisons between laboratories.",
author = "Clara Johansson and Peter M{\o}ller and Lykke Forchhammer and Steffen Loft and Godschalk, {Roger W L} and Langie, {Sabine A S} and Stijn Lumeij and Jones, {George D D} and Kwok, {Rachel W L} and Amaya Azqueta and Phillips, {David H} and Osman Sozeri and Routledge, {Michael N} and Charlton, {Alexander J} and Patrizia Riso and Marisa Porrini and Alessandra Allione and Giuseppe Matullo and Jadwiga Palus and Maciej Stepnik and Collins, {Andrew R} and Lennart M{\"o}ller",
year = "2010",
doi = "10.1093/mutage/gep055",
language = "English",
volume = "25",
pages = "125--32",
journal = "Mutagenesis",
issn = "0267-8357",
publisher = "Oxford University Press",
number = "2",

}

RIS

TY - JOUR

T1 - An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay

AU - Johansson, Clara

AU - Møller, Peter

AU - Forchhammer, Lykke

AU - Loft, Steffen

AU - Godschalk, Roger W L

AU - Langie, Sabine A S

AU - Lumeij, Stijn

AU - Jones, George D D

AU - Kwok, Rachel W L

AU - Azqueta, Amaya

AU - Phillips, David H

AU - Sozeri, Osman

AU - Routledge, Michael N

AU - Charlton, Alexander J

AU - Riso, Patrizia

AU - Porrini, Marisa

AU - Allione, Alessandra

AU - Matullo, Giuseppe

AU - Palus, Jadwiga

AU - Stepnik, Maciej

AU - Collins, Andrew R

AU - Möller, Lennart

PY - 2010

Y1 - 2010

N2 - The increasing use of single cell gel electrophoresis (the comet assay) highlights its popularity as a method for detecting DNA damage, including the use of enzymes for assessment of oxidatively damaged DNA. However, comparison of DNA damage levels between laboratories can be difficult due to differences in assay protocols (e.g. lysis conditions, enzyme treatment, the duration of the alkaline treatment and electrophoresis) and in the end points used for reporting results (e.g. %DNA in tail, arbitrary units, tail moment and tail length). One way to facilitate comparisons is to convert primary comet assay end points to number of lesions/10(6) bp by calibration with ionizing radiation. The aim of this study was to investigate the inter-laboratory variation in assessment of oxidatively damaged DNA by the comet assay in terms of oxidized purines converted to strand breaks with formamidopyrimidine DNA glycosylase (FPG). Coded samples with DNA oxidation damage induced by treatment with different concentrations of photosensitizer (Ro 19-8022) plus light and calibration samples irradiated with ionizing radiation were distributed to the 10 participating laboratories to measure DNA damage using their own comet assay protocols. Nine of 10 laboratories reported the same ranking of the level of damage in the coded samples. The variation in assessment of oxidatively damaged DNA was largely due to differences in protocols. After conversion of the data to lesions/10(6) bp using laboratory-specific calibration curves, the variation between the laboratories was reduced. The contribution of the concentration of photosensitizer to the variation in net FPG-sensitive sites increased from 49 to 73%, whereas the inter-laboratory variation decreased. The participating laboratories were successful in finding a dose-response of oxidatively damaged DNA in coded samples, but there remains a need to standardize the protocols to enable direct comparisons between laboratories.

AB - The increasing use of single cell gel electrophoresis (the comet assay) highlights its popularity as a method for detecting DNA damage, including the use of enzymes for assessment of oxidatively damaged DNA. However, comparison of DNA damage levels between laboratories can be difficult due to differences in assay protocols (e.g. lysis conditions, enzyme treatment, the duration of the alkaline treatment and electrophoresis) and in the end points used for reporting results (e.g. %DNA in tail, arbitrary units, tail moment and tail length). One way to facilitate comparisons is to convert primary comet assay end points to number of lesions/10(6) bp by calibration with ionizing radiation. The aim of this study was to investigate the inter-laboratory variation in assessment of oxidatively damaged DNA by the comet assay in terms of oxidized purines converted to strand breaks with formamidopyrimidine DNA glycosylase (FPG). Coded samples with DNA oxidation damage induced by treatment with different concentrations of photosensitizer (Ro 19-8022) plus light and calibration samples irradiated with ionizing radiation were distributed to the 10 participating laboratories to measure DNA damage using their own comet assay protocols. Nine of 10 laboratories reported the same ranking of the level of damage in the coded samples. The variation in assessment of oxidatively damaged DNA was largely due to differences in protocols. After conversion of the data to lesions/10(6) bp using laboratory-specific calibration curves, the variation between the laboratories was reduced. The contribution of the concentration of photosensitizer to the variation in net FPG-sensitive sites increased from 49 to 73%, whereas the inter-laboratory variation decreased. The participating laboratories were successful in finding a dose-response of oxidatively damaged DNA in coded samples, but there remains a need to standardize the protocols to enable direct comparisons between laboratories.

U2 - 10.1093/mutage/gep055

DO - 10.1093/mutage/gep055

M3 - Journal article

C2 - 19948595

VL - 25

SP - 125

EP - 132

JO - Mutagenesis

JF - Mutagenesis

SN - 0267-8357

IS - 2

ER -

ID: 18360051