Summary: | The emergence of carbapenem-resistant <i>Acinetobacter calcoaceticus</i>-<i>baumannii</i> complex (CRACB) in clinical environments is a significant global concern. These critical pathogens have shown resistance to a broad spectrum of antibacterial drugs, including carbapenems, mostly due to the acquisition of various β-lactamase genes. Clinical samples (n = 1985) were collected aseptically from multiple sources and grown on blood and MacConkey agar. Isolates and antimicrobial susceptibility were confirmed with the VITEK-2 system. The modified Hodge test confirmed the CRACB phenotype, and specific PCR primers were used for the molecular identification of <i>bla</i><sub>OXA</sub> and <i>bla</i><sub>NDM</sub> genes. Of the 1985 samples, 1250 (62.9%) were culture-positive and 200 (43.9%) were CRACB isolates. Of these isolates, 35.4% were recovered from pus samples and 23.5% from tracheal secretions obtained from patients in intensive care units (49.3%) and medical wards (20.2%). An antibiogram indicated that 100% of the CRACB isolates were resistant to β-lactam antibiotics and β-lactam inhibitors, 86.5% to ciprofloxacin, and 83.5% to amikacin, while the most effective antibiotics were tigecycline and colistin. The CRACB isolates displayed resistance to eight different AWaRe classes of antibiotics. All isolates exhibited the <i>bla</i><sub>OXA-51</sub> gene, while <i>bla</i><sub>OXA-23</sub> was present in 94.5%, <i>bla</i><sub>VIM</sub> in 37%, and <i>bla</i><sub>NDM</sub> in 14% of the isolates. The <i>bla</i><sub>OXA-51</sub>, <i>bla</i><sub>OXA-23</sub>, and <i>bla</i><sub>OXA-24</sub> genes co-existed in 13 (6.5%) isolates. CRACB isolates with co-existing <i>bla</i><sub>OXA-23</sub>, <i>bla</i><sub>OXA-24</sub>, <i>bla</i><sub>NDM</sub>, <i>bla</i><sub>OXA-51</sub> and <i>bla</i><sub>VIM</sub> genes were highly prevalent in clinical samples from Pakistan. CRACB strains were highly critical pathogens and presented resistance to virtually all antibacterial drugs, except tigecycline and colistin.
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