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Abstract : Despite the widespread availability of antibiotics, urinary tract infection (UTI) remains the most common bacterial infection in the human population. Current knowledge of the organisms that cause UTI and their antibiotic susceptibility pattern is mandatory to ensure appropriate therapy. Production of extended-spectrum β-lactamases (ESBLs) is an important mechanism of resistance in Enterobacteriaceae. Knowledge of their prevalence is essential to guide appropriate antibiotic therapy. Aim of the present study was to determine the frequency of ESBLs in uropathogenic Enterobacteriaceae and their antibiogram. A total of 300 uropathogenic Enterobacteriaceae isolated from a teaching hospital. Antimicrobial susceptibility testing was performed by “Kirby Bauer's” disc diffusion method according to “Clinical Laboratory Standard Institute” (CLSI) guidelines. These isolates were tested for ESBL phenotype by CLSI phenotypic confirmatory test (CLSI-PCT). It was found that 53% of the isolates were ESBL producers. E. coli (56%) and Klebsiella species (35%) were the predominant. Nitrofurantoin, piperacillin-tazobactum and amikacin were found to be the most effective antibiotics. ESBL producers were more resistant to cephalosporins, fluoroquinolones, piperacillin-tazobactum, amoxyclav and cotrimoxazole compared to non-ESBL producers. Continued antibiotic surveillance, infection control measures and an effective antibiotic policy are required to address the problem of ESBL associated infections.