Comet Assay for Genotoxicity Evaluation of Salmonella Outer Membrane Vesicles

Outer membrane vesicles (OMVs) from different Gram-negative bacteria, including Salmonella spp., have appeared as potential acellular vaccine candidates for their role in host-pathogen interaction by delivering virulence factors and modulating immune response. Given this host-cell contact, it is crucial to assess their genotoxic effects to ensure safety as vaccine candidates. The Comet assay, a sensitive and effective method for detecting single-cell DNA strand breaks, offers an effective approach for evaluating such effects. This study explores the application of the Comet assay as a highly sensitive approach to investigate the potential genotoxicity of Salmonella-derived OMVs in mammalian cells, with important implications for understanding host-pathogen interactions and cellular DNA damage, and providing a reproducible workflow for evaluating minimal DNA strand alterations in vitro as part of Salmonella-OMV safety studies.