Introduction: Healthcare-Associated Infections (HAIs) are a public health problem, and their severity increases significantly when affecting pediatric patients in the Intensive Care Unit (ICU). Objectives: To identify the main bacteria and risk factors in a pediatric ICU. Methodology: Retrospective observational cross-sectional study conducted on children admitted to the ICU of a Teaching Hospital in the city of Cascavel-PR, from January 2015 to April 2024. Results: The majority of patients were male (58.5%), and the isolated microorganisms primarily affected the respiratory tract. Of the 65 children with HAIs in the ICU, 3 (4.62%) were diagnosed with Sars-Cov-2 and 5 (7.69%) died. Among the 41 bacterial species isolated, the most prevalent were Pseudomonas aeruginosa (8; 12.3%), Klebsiella pneumoniae (6; 9.2%), and Staphylococcus aureus/Staphylococcus epidermidis/Staphylococcus haemolyticus (4; 6.2%), respectively. Conclusion: The specific microbiological profile of the pediatric ICU allows for the empirical selection of antimicrobials, improving therapeutic effects and reducing the emergence of multidrug-resistant microorganisms. This knowledge contributes to the well-being of pediatric patients, meeting the objective of identifying the main bacteria and risk factors.