Sequencing analysis of efflux pump and porin genes among Pseudomonas aeruginosa isolated from different sources

Abstract

Background: Efflux pumps are multidrug resistance mechanisms that play a crucial role in the antibiotic resistance of Pseudomonas aeruginosa. In this study, we aimed to investigate the prevalence of efflux pump genes among P. aeruginosa isolated from different sources. Methods: Forty clinical isolates that were primarily identified as P. aeruginosa were obtained from different laboratories of teaching hospitals in Baghdad/Iraq. Their identification was confirmed by using 16s rDNA as a housekeeping gene (HKG). To study different six types of efflux pump and porin genes, specific primers were used to detect mex A1, mex R, mex X, mex Y, oprD1, and oprJ1 genes followed by sequencing the amplified fragment, which was analyzed by Geneious software. Results: The results of amplification showed that all the 40 isolates (100%) were compatible with P. aeruginosa. Antibiotic sensitivity test (AST) for 14 antimicrobial agents was tested using Kirby–Bauer disc diffusion method and the result of the resistance pattern was as follows: 100% for SXT and F, 95% for TGC, 80% for CIP, 60% for LEV, 55% for FEP, 50% for AK, 47.5% for TOB, 40% for CAR, 35% for PRL, 27.5% for IMP, 25% for CAZ, 22.5% for ATM, 7.5% for COL, while the percentage of EP and porin genes among the total isolates as following: mex X 100%, 97.5% for oprD1, 95% for mexR, 92.5% for each mex Y and oprJ1, mex A 82.5%. The results explained that the most resistant six isolates in AST were resistant to 11–13 antimicrobial agents that harbored all six types of efflux pumps and porin genes and these isolates were obtained from urine and burn specimens. Conclusion: our study demonstrated a high prevalence of efflux pumps and porin genes among P. aeruginosa isolates from different sources, indicating the significant role of efflux pumps in antibiotic resistance. These findings highlight the need for continuous surveillance of antibiotic resistance mechanisms and the development of new strategies to combat MDR P. aeruginosa infections.