| Summary: | Wound infections are common medical problems in sub-Saharan Africa but data on the molecular epidemiology are rare. Within this study we assessed the clonal lineages, resistance genes and virulence factors of Gram-negative bacteria isolated from Ghanaian patients with chronic wounds. From a previous study, 49 <i>Pseudomonas aeruginosa</i>, 21 <i>Klebsiella</i> <i>pneumoniae</i> complex members and 12 <i>Escherichia coli</i> were subjected to whole genome sequencing. Sequence analysis indicated high clonal diversity with only nine <i>P. aeruginosa</i> clusters comprising two strains each and one <i>E. coli</i> cluster comprising three strains with high phylogenetic relationship suggesting nosocomial transmission. Acquired beta-lactamase genes were observed in some isolates next to a broad spectrum of additional genetic resistance determinants. Phenotypical expression of extended-spectrum beta-lactamase activity in the Enterobacterales was associated with <i>bla</i><sub>CTX-M-15</sub> genes, which are frequent in Ghana. Frequently recorded virulence genes comprised genes related to invasion and iron-uptake in <i>E. coli</i>, genes related to adherence, iron-uptake, secretion systems and antiphagocytosis in <i>P. aeruginosa</i> and genes related to adherence, biofilm formation, immune evasion, iron-uptake and secretion systems in <i>K. pneumonia </i>complex. In summary, the study provides a piece in the puzzle of the molecular epidemiology of Gram-negative bacteria in chronic wounds in rural Ghana.
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