Objective: Healthcare-associated infections are critical medical conditions as they prolong hospital stays, cause multi-or- gan dysfunction, and increase mortality because of limited treatment options. In this study, we aimed to investigate the type of infection, causative agents, and resistance profiles in patients followed by healthcare-associated infections. Methods: A total of 1.235 patients diagnosed with healthcare-associated infections in our hospital between January 2016 and December 2020 were included in this study. Results: The rates of healthcare-associated infections were 0.60% in 2016, 0.27% in 2017, 0.20% in 2018, 0.89% in 2019, and 3.77% in 2020. The most common type of healthcare-associated infection was bloodstream infection in all years. We observed that healthcare-associated infections were detected more frequently in the Anesthesiology and Reanima- tion Intensive Care Unit compared to other clinics (53.3%), and Acinetobacter spp. was the most common cause with a rate of 45.2%. Gram-negative bacteria were the most frequently isolated etiologic subgroup in all years. In terms of antimicrobial resistance, while an increase in carbapenem resistance was observed in Acinetobacter spp. and Klebsi- ella pneumoniae over the years, no significant difference was found in the extended spectrum beta-lactamase (ESBL) rate in Escherichia coli. In addition, while ESBL rates decreased in Pseudomonas spp. over the years, an increase was detected in carbapenem resistance in 2017 and 2018, and we observed that the resistance remained at the same level in the following years. Conclusion: Healthcare-associated infections continue to be a significant health problem. Although our rates of healthcare-associated infections were lower than the literature, we observed that drug resistance, especially carbapenem resistance, increased in almost all agents. Acinetobacter species remained the most problematic agent.