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Bacterial resistance to antibiotics is a major public health issue, also affecting the paediatric population and leading to significant morbidity and mortality, as well as increasing healthcare costs. Overconsumption of antibiotics and prolonged hospital stays favor the emergence of these bacteria. The aim of this study was to draw up an epidemiological and bacteriological profile of ESKAPEE group multidrug-resistant bacteria isolated from clinical samples taken from patients hospitalized in the pediatric wards of a university hospital in central Tunisia, and to reveal the characteristics of carrier patients. The highest rate was observed in the pediatric cardiovascular and thoracic surgery department, reaching fifty percent. Multi-resistant Enterobacteriaceae, notably Escherichia coli resistant to third-generation cephalosporins, were the most frequently identified, particularly in urinary tract infections. The study also analyzes risk factors, treatment options and suggests prevention strategies.