DNA-repair measurements by use of the modified comet assay: an inter-laboratory comparison within the European Comet Assay Validation Group (ECVAG)

Research output: Contribution to journalJournal article

  • Roger W L Godschalk
  • Clara Ersson
  • Patrizia Riso
  • Marisa Porrini
  • Sabine A S Langie
  • Frederik-Jan van Schooten
  • Amaya Azqueta
  • Andrew R Collins
  • George D D Jones
  • Rachel W L Kwok
  • David H Phillips
  • Osman Sozeri
  • Alessandra Allione
  • Giuseppe Matullo
  • Lennart Möller
  • Lykke Forchhammer
  • Loft, Steffen
  • Møller, Peter
The measurement of DNA-repair activity by extracts from cells or tissues by means of the single-cell gel electrophoresis (comet) assay has a high potential to become widely used in biomonitoring studies. We assessed the inter-laboratory variation in reported values of DNA-repair activity on substrate cells that had been incubated with Ro19-8022 plus light to generate oxidatively damaged DNA. Eight laboratories assessed the DNA-repair activity of three cell lines (i.e. one epithelial and two fibroblast cell lines), starting with cell pellets or with cell extracts provided by the coordinating laboratory. There was a large inter-laboratory variation, as evidenced by the range in the mean level of repair incisions between the laboratory with the lowest (0.002incisions/10(6)bp) and highest (0.988incisions/10(6)bp) incision activity. Nevertheless, six out of eight laboratories reported the same cell line as having the highest level of DNA-repair activity. The two laboratories that reported discordant results (with another cell line having the highest level of DNA-repair activity) were those that reported to have little experience with the modified comet assay to assess DNA repair. The laboratories were also less consistent in ordering the repair activity of the other two cell lines, probably because the DNA-repair activity by extracts from these cell lines were very similar (on average approximately 60-65% of the cell line with the highest repair capacity). A significant correlation was observed between the repair activity found in the provided and the self-made cell extracts (r=0.71, P
Original languageEnglish
JournalMutation Research - Genetic Toxicology and Environmental Mutagenesis
Volume757
Issue number1
Pages (from-to)60-7
Number of pages8
ISSN1383-5718
DOIs
Publication statusPublished - 18 Sep 2013

    Research areas

  • Cell Line, Comet Assay, DNA Damage, DNA Repair, Environmental Monitoring, Humans, Monocytes

ID: 98468943