AUTHOR=Füszl Astrid , Pietzka Ariane , Hyden Patrick , Mösenbacher Tobias , Stöger Anna , Blaschitz Marion , Stadlbauer Silke , Hasenberger Petra , Schindler Stefanie , Heger Florian , Pleininger Sonja , Indra Alexander TITLE=Assessing mutation accumulation in DNA repair-deficient Listeria monocytogenes: implications for cgMLST cluster thresholds in outbreak analysis JOURNAL=Frontiers in Cellular and Infection Microbiology VOLUME=Volume 15 - 2025 YEAR=2025 URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2025.1530851 DOI=10.3389/fcimb.2025.1530851 ISSN=2235-2988 ABSTRACT=BackgroundListeria (L.) monocytogenes is primarily transmitted via contaminated food and can cause listeriosis, an infection often associated with sepsis and meningitis in at-risk individuals. Accurate outbreak detection relies on whole genome sequencing (WGS) and core genome multilocus sequence typing (cgMLST), which use allele thresholds to identify related strains.MethodsThis study investigated mutation rates in L. monocytogenes, focusing on isolates with DNA repair deficiencies. Serial subcultivations were performed, comparing a repair-deficient isolate with a wild-type control. Genetic variability was assessed using WGS and cgMLST.ResultsMutation rates were significantly higher in repair-deficient isolates, exceeding typical cgMLST thresholds currently used in Listeria outbreak investigations, leading to a misclassification of related isolates as unrelated. An additional analysis of the Austrian Listeria database revealed that such deficiencies are rare among isolates.ConclusionsThe standard 7-allele cgMLST threshold effectively identifies related strains in most cases, but may require adjustments for hypermutator strains. Incorporating DNA repair data could improve the accuracy of outbreak investigations, ensuring reliable public health responses.