In recent years, healthcare-associated infections (HAIs) have been a priority topic for global institutions such as the World Health Organization because, during the COVID-19 pandemic, the role of HAIs as co-morbidity in infected patients was evident. HAIs can cause disability and mortality and lead to excessive healthcare costs. This work aims to calculate the prevalence of HAIs from 2019 to 2023 to determine their microbiological profile and antibiotic resistance. A cross-sectional design was used for this study. To describe the population, univariate analysis, measures of central tendency, frequencies, and proportions were used. The present study included 3,936 HAI notifications, which showed a significant decrease in their number during the COVID-19 pandemic, especially in 2020. In 2021 and 2022, the numbers increased rapidly to around 30%. The most prevalent HAI type was ventilator-associated pneumonia, which had the highest prevalence in 2020. Regarding microorganism isolation, percentages increased after 2020 from 46% to 67%, with Acinetobacter baumannniibeingthe most frequent during and after pandemics. The microbiological profile showed multidrug resistance in several microorganisms. HAIs are a global health concern. The number of cases has been increasing since the COVID-19 pandemic with regard to the multidrug-resistant microorganism. HAIs have an important impact on the quality of life, morbidity, mortality, and financial concerns for health services. Global strategies should be adapted for different regions, since the panaroma in developed countries allows for programs to be established for the prevention and control of HAIs in a better way than in low-income countries that lack adequate infrastructure and resources.