Antimicrobial resistance (AMR) poses a global threat, with carbapenem-resistant Enterobacterales (CRE) representing a significant concern due to limited therapeutic options. This study investigated the prevalence of carbapenemase genes in CRE strains isolated from tracheal aspirates of patients at a Brazilian university hospital between January 2020 and August 2023. Bacterial identification was conducted using MALDI-TOF, while carbapenemase genes were detected by qPCR. Demographic and clinical data were collected, and univariate analysis was performed using the chi-square test (p < 0.05). Variables with p ≤ 0.10 were further investigated using the chi-square test for linear trend, along with stratified analysis. Out of 1,133 samples, 111 (9.79%) showed CRE growth, with 46 isolates included in the final sample, predominantly comprising Klebsiella pneumoniae (65.21%) and Serratia marcescens (19.57%). The blaKPC gene was prevalent (78.26%), while blaNDM was detected in 21.74% of cases. The identified population was predominantly male (67.39%), elderly (69.57%), white (56.52%), unmarried (63.04%), and had a low level of education (56.52%). Most patients (69.57%) were in the intensive care unit and remained hospitalized for more than 30 days (76.08%). There was a significant inverse trend between Klebsiella pneumoniae and age (p = 0.045), as well as a direct linear trend between blaNDM and the annual increase in COVID-19 cases in Brazil (p = 0.050). A high probability of finding non-Klebsiella pneumoniae bacteria was observed in patients with prolonged hospital stays, independent of COVID-19 (p = 0.006) and the type of resistance genes (p = 0.020). The persistent prevalence of CRE, especially with blaKPC, underscores the urgency of effective control measures.
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