This study assessed the diagnostic performance of the VITEK®2 system in identifying bacterial pathogens from 150 clinical specimens (blood, urine, and stool) obtained from patients aged 10–70 years. Bacterial isolates were characterized morphologically and biochemically, followed by identification and antimicrobial susceptibility testing (AST) using the VITEK®2 system. Nine pathogenic bacteria were isolated, including Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Salmonella typhi, and Bacillus species. The VITEK®2 system demonstrated high confidence in identification (90–99% probability), with excellent performance for common pathogens like E. coli and S. typhi. Direct Susceptibility Testing (DST) revealed alarming resistance patterns, particularly to beta-lactam antibiotics (ampicillin, cefotaxime) in Gram-negative isolates, likely due to extended-spectrum beta-lactamase (ESBL) production. Resistance to azithromycin in P. aeruginosa and Enterobacter aerogenes was linked to efflux pumps or ribosomal modifications, while fluoroquinolone resistance in S. typhi suggested mutations in DNA gyrase/topoisomerase IV. Notably, multidrug-resistant (MDR) strains, such as P. aeruginosa (resistant to ampicillin, cefotaxime, and azithromycin), were identified, underscoring the urgency of antimicrobial stewardship. The VITEK®2 system rapidly provided accurate results within 3 hours, improving treatment decisions, but the high resistance rates highlight the need for stronger surveillance and alternative strategies such as phage or combination therapies

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