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Title: | Potassium bromate as positive assay control for the Fpg-modified comet assay | Authors: | Moller, Peter Muruzabal, Damian Bakuradze, Tamara Richling, Elke Bankoglu, Ezgi Eyluel Stopper, Helga LANGIE, Sabine Azqueta, Amaya Jensen, Annie Scavone, Francesca Giovannelli, Lisa Wojewodzka, Maria Kruszewski, Marcin Valdiglesias, Vanessa Laffon, Blanca Costa, Carla Costa, Solange Teixeira, Joao Paulo Marino, Mirko Del Bo', Cristian Riso, Patrizia Shaposhnikov, Sergey Collins, Andrew |
Issue Date: | 2020 | Publisher: | OXFORD UNIV PRESS | Source: | MUTAGENESIS, 35 (4) , p. 341 -347 | Abstract: | The comet assay is a popular assay in biomonitoring studies. DNA strand breaks (or unspecific DNA lesions) are measured using the standard comet assay. Oxidative stress-generated DNA lesions can be measured by employing DNA repair enzymes to recognise oxidatively damaged DNA. Unfortunately, there has been a tendency to fail to report results from assay controls (or maybe even not to employ assay controls). We believe this might have been due to uncertainty as to what really constitutes a positive control. It should go without saying that a biomonitoring study cannot have a positive control group as it is unethical to expose healthy humans to DNA damaging (and thus potentially carcinogenic) agents. However, it is possible to include assay controls in the analysis (here meant as a cryopreserved sample of cells i.e. included in each experiment as a reference sample). In the present report we tested potassium bromate (KBrO3) as a positive comet assay control for the formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay. Ten laboratories used the same procedure for treatment of monocytic THP-1 cells with KBrO3 (0.5, 1.5 and 4.5 mM for 1 h at 37 degrees C) and subsequent cryopreservation. Results from one laboratory were excluded in the statistical analysis because of technical issues in the Fpg-modified comet assay. All other laboratories found a concentration-response relationship in cryopreserved samples (regression coefficients from 0.80 to 0.98), although with different slopes ranging from 1.25 to 11.9 Fpg-sensitive sites (%DNA in tail) per 1 mM KBrO3. Our results demonstrate that KBrO3 is a suitable positive comet assay control. | Notes: | Moller, P (corresponding author), Univ Copenhagen, Dept Publ Hlth, Sect Environm Hlth, Oster Farimagsgade 5A, DK-1014 Copenhagen K, Denmark. pemo@sund.ku.dk |
Other: | Moller, P (corresponding author), Univ Copenhagen, Dept Publ Hlth, Sect Environm Hlth, Oster Farimagsgade 5A, DK-1014 Copenhagen K, Denmark. pemo@sund.ku.dk | Keywords: | potassium;bromates;dna;laboratory;biological monitoring;positive controlt;hp-1 cells;comet assay;cryopreservation | Document URI: | http://hdl.handle.net/1942/33360 | ISSN: | 0267-8357 | e-ISSN: | 1464-3804 | DOI: | 10.1093/mutage/geaa011 | ISI #: | WOS:000606985200005 | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2022 |
Appears in Collections: | Research publications |
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