Acinetobacter baumannii is a significant nosocomial pathogen recognized for its multidrug-resistance (MDR) and capacity to endure in hospital settings. This study aims to investigate the clonal relationships of A. baumannii isolates from diverse clinical samples, identify the sequence types of MDR isolates, and examine biofilm formation activity and biofilm-associated genes that contribute to persistence in hospital settings. A total of 90 A. baumannii isolates were analyzed. Bacterial identification and antibiotic susceptibility testing were conducted with MALDI-TOF MS and Vitek-2. REP-PCR was utilized to evaluate clonal connections, MLST was employed for specific isolates. Biofilm formation activity was assessed using the XTT reduction assay, and biofilm-associated genes were identified by PCR. REP-PCR revealed 29 genotypes, with Genotype A being identified as the endemic clone in 59% of isolates. Two isolates representing this genotype were found to belong to the ST2 clone. The majority of A. baumannii isolates possess biofilm-related genes and exhibit strong biofilm activity. In MDR isolates, ompA and csuE positivity were significantly higher than those non-MDR isolates (P = 0.003, P = 0.001). The csuE positive isolates were found to have significantly stronger biofilm activity than negative ones (P = 0.009). This study emphasizes the prevalence of a hospital-endemic, MDR A. baumannii genotype A, ST2 clone, and the genetic variability across isolates. No direct correlation was noted between MDR status and biofilm formation; however, some biofilm-related genes, notably csuE, were linked to stronger biofilm activity. These findings underscore the necessity for ongoing molecular surveillance and infection control measures to avert the dissemination of MDR A. baumannii in healthcare environments.
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