Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories

Roger W L Godschalk, Clara Ersson, Maciej Stępnik, Magdalena Ferlińska, Jadwiga Palus, João Paulo Teixeira, Solange Costa, George D D Jones, Jennifer A. Higgins, Johanna Kain, Lennart Möller, Lykke Forchhammer, Steffen Loft, Yolanda Lorenzo, Andrew R. Collins, Frederik Jan van Schooten, Blanca Laffon, Vanessa Valdiglesias, Marcus Cooke, Vilas MistryMahsa Karbaschi, David H. Phillips, Osman Sozeri, Michael N. Routledge, Kirsty Nelson-Smith, Patrizia Riso, Marisa Porrini, Adela López de Cerain, Amaya Azqueta, Giuseppe Matullo, Alessandra Allione, Peter Møller

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Abstract

This study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp).

Original languageEnglish
Pages (from-to)241-249
Number of pages9
JournalMutagenesis
Volume29
Issue number4
DOIs
Publication statusPublished - Jul 1 2014

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ASJC Scopus subject areas

  • Medicine(all)

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Godschalk, R. W. L., Ersson, C., Stępnik, M., Ferlińska, M., Palus, J., Teixeira, J. P., Costa, S., Jones, G. D. D., Higgins, J. A., Kain, J., Möller, L., Forchhammer, L., Loft, S., Lorenzo, Y., Collins, A. R., van Schooten, F. J., Laffon, B., Valdiglesias, V., Cooke, M., ... Møller, P. (2014). Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories. Mutagenesis, 29(4), 241-249. https://doi.org/10.1093/mutage/geu012