Quantifying the expression levels of mexA and mexB genes in response to the efflux pump inhibitor PAβN in ciprofloxacin-resistant Pseudomonas aeruginosa isolated from urinary tract infections
Abstract
Urinary tract infections (UTIs) are among the most prevalent bacterial infections worldwide, occurring in both community and healthcare settings. Pseudomonas aeruginosa, a Gram-negative opportunistic pathogen, is one of the five most significant nosocomial bacteria and a major contributor to UTIs. This study aimed to quantify the expression levels of efflux pump genes mexA and mexB in response to the efflux pump inhibitor phenylalanine-arginine β-naphthylamide (PAβN) in ciprofloxacin-resistant P. aeruginosa isolates.
Methods. Fifty urine specimens were collected from UTI patients at various hospitals in Baghdad. Specimens were directly cultured by streaking on differential media. Five ciprofloxacin-resistant P. aeruginosa isolates were identified, with resistance confirmed using the disk diffusion method for antibiotic susceptibility. The broth microdilution method was employed to determine the minimum inhibitory concentration (MIC) of ciprofloxacin (CIP) alone and in combination with PAβN to assess PAβN’s inhibitory activity. RNA was extracted and purified from the bacterial isolates, followed by reverse transcription and quantitative PCR to evaluate the expression of efflux pump-related genes. The expression levels of mexA and mexB were measured in the presence of the tested compounds using quantitative PCR.
Results. Antibiotic susceptibility testing revealed that the isolates were resistant to nearly all antibiotics tested, except piperacillin-tazobactam, which was…
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