Abstract

Background: Acinetobacter baumannii is an important opportunistic pathogen responsible for healthcare-associated infections, particularly among critically ill, intensive care unit (ICU), and post-surgical patients. The emergence of carbapenem-resistant A. baumannii (CRAB) has become a major therapeutic challenge worldwide because of its multidrug-resistant nature and limited treatment options. Despite the increasing prevalence of CRAB in Pakistan, information regarding the molecular characterization of carbapenem resistance genes among isolates recovered from surgical site infections (SSIs) remains limited. Methods: A cross-sectional study was conducted from November 2023 to February 2024 at Khalifa Gul Nawaz Hospital, Bannu, Khyber Pakhtunkhwa, Pakistan. A total of (n = 118) surgical wound specimens were collected from patients with clinically diagnosed SSIs. A. baumannii isolates were identified using standard biochemical tests and confirmed by 16S rRNA gene sequencing. In vitro antimicrobial susceptibility was determined by Kirby–Bauer disk diffusion according to CLSI 2024 guidelines. In addition, the concentration-dependent inhibition-zone response of imipenem and meropenem against carbapenem-resistant isolates was evaluated using an agar well diffusion assay. Molecular detection of blaNDM-1 and blaOXA-23 was performed by polymerase chain reaction (PCR), and representative amplicons were subjected to Sanger sequencing for further analysis. Results: In the current study, the total number of (n = 118) wound specimens from healthcare-associated patients were processed, in which A. baumannii-positive isolates (n = 23, 19.5%) were documented. Its prevalence was higher in males (69.6%) compared to females, and was strongly associated (78.2%) with elderly patients (aged 51–68 years). In vitro susceptibility testing revealed that 91.3% of isolates harbored multidrug-resistant (MDR) attributes. Antimicrobial susceptibility profiling revealed high levels of multidrug resistance, with 82.6% of isolates resistant to imipenem and meropenem, as well as 100% resistance to aztreonam and gentamicin. Colistin showed the highest activity, with 87.0% of isolates remaining susceptible. In the agar well diffusion assay, measurable inhibition of the carbapenem-resistant isolates by imipenem and meropenem occurred only at higher concentrations. Molecular screening of resistance genes identified blaOXA-23 in 60.9% and blaNDM-1 in 30.4% of isolates. Co-expression of both genes was detected in some (n = 4) isolates. Among the selected MDR isolates included in the gene resistance analysis, imipenem and meropenem resistance were recorded in 9/14 blaOXA-23-positive isolates and 5/7 blaNDM-1-positive isolates. Conclusions: This study substantiates an escalated incidence of carbapenem-resistant, MDR A. baumannii in post-surgical infections. blaOXA-23 were documented as the predominant carbapenemase gene, with co-expression of blaNDM-1 in a substantial proportion of isolates. These findings provide an important phenotypic and molecular characterization of selected carbapenemase genes in healthcare-associated infections, highlighting the exigency for strengthened antimicrobial stewardship and infection control strategies in the hospitals of the region.

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