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  • Title: Enhancement of genotoxic effects in the comet assay with human blood samples by aphidicolin.
    Author: Speit G, Schütz P, Hoffmann H.
    Journal: Toxicol Lett; 2004 Nov 28; 153(3):303-10. PubMed ID: 15454306.
    Abstract:
    The comet assay (single cell gel electrophoresis) has become increasingly used in human biomonitoring. In its standard version at pH > 13, DNA lesions such as DNA double-strand breaks (DSB), DNA single strand breaks (SSB) and alkali-labile sites (ALS) lead to increased DNA migration. Besides DNA damage, strand break formation during excision repair can also increase DNA migration. Inhibitors of DNA repair have been shown to enhance the DNA effects of mutagens and the use of repair inhibitors has been proposed for human biomonitoring studies to increase the sensitivity of the comet assay. To further evaluate the usefulness of such an approach we performed an experimental study with human blood and tested the enhancing effect of aphidicolin (APC) on DNA effects induced by different mutagens. Our results clearly show that APC enhances the genotoxic effects of benzo[a]pyrene diolepoxide (BPDE), bischloroethylnitrosurea (BCNU) and methyl methanesulfonate (MMS), but has no significant effect on gamma radiation-induced DNA effects. The enhancing effect is seen in unstimulated and PHA-stimulated blood, indicating repair activity under both conditions but the effect is stronger in stimulated blood. Our results indicate that APC can be used to increase the sensitivity of the comet assay towards a broad spectrum of induced primary DNA lesions and support the usefulness of this approach. However, for human biomonitoring, a sensitive protocol still has to be established.
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