Detection of carbapenem resistance mechanisms in difficult-to-treat Pseudomonas aeruginosa clinical isolates from a tertiary care hospital in Egypt

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Salem D.; Zaiton M.; El-Shenawy A.; Dahroug H.; El-Dein S.; Gamal D.; Althoqapy A.A.; Mohamed A.A.W.; Shemis M.A.; Diab M.; Maklad S.S.

Journal: BMC Microbiology

Publisher: BioMed Central Ltd

Publication Date: 4 September 2026

Volume / Issue: Volume 26 / Issue 1

Article No.: 785

ISSN: 14712180

DOI: 10.1186/s12866-026-05550-2

Scopus: View on Scopus

PubMed: 42693412

Document Type: Article

Access: All Open Access; Gold Open Access; Green Open Access

Authors and Affiliations

Salem D., Microbiology Department, Theodor Bilharz Research Institute (TBRI), P.O box 12411, 30 Imbaba, Giza, Egypt; Zaiton M., Microbiology Department, Theodor Bilharz Research Institute (TBRI), P.O box 12411, 30 Imbaba, Giza, Egypt; El-Shenawy A., Microbiology Department, Theodor Bilharz Research Institute (TBRI), P.O box 12411, 30 Imbaba, Giza, Egypt; Dahroug H., Microbiology Department, Theodor Bilharz Research Institute (TBRI), P.O box 12411, 30 Imbaba, Giza, Egypt; El-Dein S., Microbiology Department, Theodor Bilharz Research Institute (TBRI), P.O box 12411, 30 Imbaba, Giza, Egypt; Gamal D., Microbiology Department, Theodor Bilharz Research Institute (TBRI), P.O box 12411, 30 Imbaba, Giza, Egypt; Althoqapy A.A., Medical Microbiology and Immunology, Faculty of Medicine for Girls, Al- Azhar University, Cairo, Egypt; Mohamed A.A.W., Biochemistry and Molecular Biology Department, Theodor Bilharz Research institute (TBRI), P.O box 12411, 30 Imbaba, Giza, Egypt; Shemis M.A., Biochemistry and Molecular Biology Department, Theodor Bilharz Research institute (TBRI), P.O box 12411, 30 Imbaba, Giza, Egypt; Diab M., Microbiology Department, Theodor Bilharz Research Institute (TBRI), P.O box 12411, 30 Imbaba, Giza, Egypt; Maklad S.S., Medical Microbiology and Immunology, Faculty of Medicine for Girls, Al- Azhar University, Cairo, Egypt

Abstract

Background: The increasing incidence of carbapenem-resistant Pseudomonas aeruginosa (CRPA) poses a significant global health concern, particularly with the emergence of difficult-to-treat resistance (DTR), which seriously restricts therapeutic options. Therefore, we aimed to characterize DTR-PA isolates among the studied CRPA; regarding their resistance mechanisms to carbapenems and their antimicrobial susceptibility profiles. Methods: Fifty-two non-duplicate CRPA isolates were collected from clinical specimens. Antimicrobial susceptibility testing was performed using disc diffusion and VITEK2 compact system. Isolates were classified as multidrug-resistant P. aeruginosa (MDR-PA), extensively drug-resistant P. aeruginosa (XDR-PA), and DTR-PA. Conventional PCR was used to detect carbapenemase genes (blaNDM, blaVIM, blaIMP, blaKPC, and blaOXA-48). Quantitative real-time PCR (qRT-PCR) identified the over expression of efflux pump genes (mexB, mexY) and the down regulation of outer membrane porin (oprD) gene. Results: Fifty-two (52) CRPA isolates were categorized according to resistance phenotype into three main groups where 46% were identified as DTR-PA, 38% as XDR-PA, 8% as MDR-PA and 8% remained uncategorized. In DTR-PA, carbapenemase genes were detected in 92% of isolates with blaNDM being the most prevalent (75%), followed by blaVIM (54%) and blaOXA-48 (38%). Co-existence of multiple carbapenemase genes was common. Efflux pump mechanism was the most common mechanism of resistance as overexpression of MexB and MexY was observed in 98% and 71% of isolates, respectively, whereas downregulation of oprD was detected in 92%. The DTR-PA phenotype demonstrated the least sensitive profiles and most of the isolates (92%) were positive to all the studied resistant mechanisms (carbapenemase production, overexpression of efflux pump, and porin loss) with a statistically significant association (P = 0.001). Conclusions: Difficult to treat Pseudomonas aeruginosa is highly prevalent among CRPA isolates in our setting and is caused by multiple resistance mechanisms, particularly blaNDM carriage, efflux pump overexpression, and oprD downregulation. These findings underscore the urgent need for enhanced molecular surveillance and antimicrobial stewardship to limit further dissemination of highly resistant Pseudomonas aeruginosa (P. aeruginosa) iaolates. Future studies utilizing the DTR-PA definition are needed, as this category highlights the most resistant CRPA isolates with maximum resistance profiles that most likely to pose clinical challenges. © The Author(s) 2026.

Keywords

bla<sub>NDM</sub>; Carbapenem resistant Pseudomonas aeruginosa (CRPA); Carbapenemases; Difficult-to-treat resistance (DTR); Efflux pumps; MexB; MexY; Multidrug resistance (MDR); OprD; Anti-Bacterial Agents; Bacterial Outer Membrane Proteins; Bacterial Proteins; beta-Lactamases; Carbapenems; Drug Resistance, Multiple, Bacterial; Egypt; Humans; Membrane Transport Proteins; Microbial Sensitivity Tests; Porins; Pseudomonas aeruginosa; Pseudomonas Infections; Tertiary Care Centers; amikacin; aminoglycoside; aztreonam; carbapenemase; cefepime; ceftazidime; imipenem; meropenem; piperacillin plus tazobactam; porin; tobramycin; antiinfective agent; bacterial protein; beta lactamase; carbapenem derivative; carrier protein; OprD protein, Pseudomonas aeruginosa; outer membrane protein; antibiotic sensitivity; Article; bacterial gene; bacterium isolation; carbapenem resistance; cohort analysis; controlled study; diagnostic test accuracy study; disk diffusion; DNA extraction; human; mexB gene; mexY gene; multidrug resistance; oprD gene; polymerase chain reaction; predictive value; quantitative analysis; sensitivity and specificity; tertiary health care; drug effect; drug therapy; genetics; isolation and purification; microbial sensitivity test; microbiology; Pseudomonas infection; tertiary care center

Citation Information

Scopus Citations: 0

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