Diagnostic algorithm for the detection of carbapenemases and extended-spectrum β-lactamases in carbapenem-resistant Pseudomonas aeruginosa
ABSTRACT Pseudomonas aeruginosa can acquire carbapenem resistance through various mechanisms, including genomic mutations leading to the overexpression of efflux pumps, intrinsic AmpC-β-lactamase, and/or reduced permeability, and/or through the acquisition of plasmid-mediated carbapenemases and/or e...
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| Language: | English |
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American Society for Microbiology
2025-06-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.03196-24 |
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| author | Stefano Mancini Laia Garcia-Verellen Helena M. B. Seth-Smith Peter M. Keller Natalia Kolesnik-Goldmann Muhammad Ali Syed Irfan Ullah Vladimira Hinic Tim Roloff Adrian Egli Oliver Nolte |
| author_facet | Stefano Mancini Laia Garcia-Verellen Helena M. B. Seth-Smith Peter M. Keller Natalia Kolesnik-Goldmann Muhammad Ali Syed Irfan Ullah Vladimira Hinic Tim Roloff Adrian Egli Oliver Nolte |
| author_sort | Stefano Mancini |
| collection | DOAJ |
| description | ABSTRACT Pseudomonas aeruginosa can acquire carbapenem resistance through various mechanisms, including genomic mutations leading to the overexpression of efflux pumps, intrinsic AmpC-β-lactamase, and/or reduced permeability, and/or through the acquisition of plasmid-mediated carbapenemases and/or extended-spectrum-β-lactamases (ESBLs). Unfortunately, carbapenemase/ESBL-producing-carbapenem-resistant-P. aeruginosa (CP/ESBL-CRPA) cannot be differentiated from non-CP/ESBL-CRPA based solely on susceptibility testing results of conventional β-lactam (BL)-antibiotics. Knowing that these two groups display different activity profiles toward novel BL/β-lactamase-inhibitor (BLI) combinations, we developed and verified a cost-effective and easy-to-use diagnostic algorithm for screening and differentiation of carbapenemase and ESBL production in CRPA. We determined disc diffusion inhibition zones and gradient strip minimal inhibitory concentration values of 136 whole-genome-sequenced CRPA (70 metallo-β-lactamase-[MBL-], 1 GES-5-, 1 KPC-2-, 12 ESBL-, and 53 AmpC-hyper-producing isolates). We used the following BL-BLI combinations: ceftolozane-tazobactam (C-T), ceftazidime-avibactam, imipenem-relebactam (I-R), meropenem-vaborbactam, cefepime-enmetazobactam (C-E), and aztreonam-avibactam. We also included a lateral flow immunoassay (Carba-5, NG-Biotech) for confirmation of MBL production and double disc synergy testing (DDST) to improve ESBL detection. C-T was the most effective screening antibiotic for distinguishing MBL and ESBL producers from AmpC-hyperproducing CRPA, achieving a sensitivity of 100% for both MBL and ESBL producers. I-R reliably confirmed MBL production in C-T positive screened CRPAs, with a sensitivity of 92.8% and specificity of 100%. Incorporating Carba-5 into the phenotypic algorithm improved sensitivity for confirming MBL production to 100%. For the remaining C-T positive but I-R negative isolates, C-E showed 75% sensitivity and 78.6% specificity in detecting ESBL production. The DDST further confirmed ESBL production in six out of nine ESBL producers (66.6%). In conclusion, we established a simple and cost-effective diagnostic algorithm, enabling screening and confirmation of carbapenemase and ESBL production in CRPA.IMPORTANCECarbapenem-resistant Pseudomonas aeruginosa (CRPA) is a major global health threat, and rapid identification of its resistance mechanisms is crucial for effective treatment and infection control. Differentiating between carbapenemase-producing (CP), extended-spectrum β-lactamase-producing (ESBL), and AmpC-hyperproducing CRPA is challenging, as conventional susceptibility testing cannot reliably distinguish these resistance mechanisms. Our study presents a simple, cost-effective, and easy-to-implement phenotypic diagnostic algorithm that enables accurate screening and confirmation of CP and ESBL production in CRPA. This method is particularly valuable for laboratories lacking access to molecular diagnostics, as it provides a practical alternative for routine testing. By facilitating the early detection of resistant P. aeruginosa strains, this approach has the potential to improve patient outcomes, optimize antimicrobial therapy, and enhance global surveillance efforts against multidrug-resistant pathogens. |
| format | Article |
| id | doaj-art-3214fd482e0945b5b128b90daaa84b81 |
| institution | OA Journals |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-3214fd482e0945b5b128b90daaa84b812025-08-20T02:05:28ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-06-0113610.1128/spectrum.03196-24Diagnostic algorithm for the detection of carbapenemases and extended-spectrum β-lactamases in carbapenem-resistant Pseudomonas aeruginosaStefano Mancini0Laia Garcia-Verellen1Helena M. B. Seth-Smith2Peter M. Keller3Natalia Kolesnik-Goldmann4Muhammad Ali Syed5Irfan Ullah6Vladimira Hinic7Tim Roloff8Adrian Egli9Oliver Nolte10Institute of Medical Microbiology, University of Zurich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zurich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zurich, Zürich, SwitzerlandUniversity Hospital Basel, University of Basel, Basel, SwitzerlandUniversity Hospital Basel, University of Basel, Basel, SwitzerlandDepartment of Microbiology, The University of Haripur, Haripur, Khyber Pakhtunkhwa, PakistanDepartment of Microbiology, The University of Haripur, Haripur, Khyber Pakhtunkhwa, PakistanInstitute of Medical Microbiology, University of Zurich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zurich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zurich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zurich, Zürich, SwitzerlandABSTRACT Pseudomonas aeruginosa can acquire carbapenem resistance through various mechanisms, including genomic mutations leading to the overexpression of efflux pumps, intrinsic AmpC-β-lactamase, and/or reduced permeability, and/or through the acquisition of plasmid-mediated carbapenemases and/or extended-spectrum-β-lactamases (ESBLs). Unfortunately, carbapenemase/ESBL-producing-carbapenem-resistant-P. aeruginosa (CP/ESBL-CRPA) cannot be differentiated from non-CP/ESBL-CRPA based solely on susceptibility testing results of conventional β-lactam (BL)-antibiotics. Knowing that these two groups display different activity profiles toward novel BL/β-lactamase-inhibitor (BLI) combinations, we developed and verified a cost-effective and easy-to-use diagnostic algorithm for screening and differentiation of carbapenemase and ESBL production in CRPA. We determined disc diffusion inhibition zones and gradient strip minimal inhibitory concentration values of 136 whole-genome-sequenced CRPA (70 metallo-β-lactamase-[MBL-], 1 GES-5-, 1 KPC-2-, 12 ESBL-, and 53 AmpC-hyper-producing isolates). We used the following BL-BLI combinations: ceftolozane-tazobactam (C-T), ceftazidime-avibactam, imipenem-relebactam (I-R), meropenem-vaborbactam, cefepime-enmetazobactam (C-E), and aztreonam-avibactam. We also included a lateral flow immunoassay (Carba-5, NG-Biotech) for confirmation of MBL production and double disc synergy testing (DDST) to improve ESBL detection. C-T was the most effective screening antibiotic for distinguishing MBL and ESBL producers from AmpC-hyperproducing CRPA, achieving a sensitivity of 100% for both MBL and ESBL producers. I-R reliably confirmed MBL production in C-T positive screened CRPAs, with a sensitivity of 92.8% and specificity of 100%. Incorporating Carba-5 into the phenotypic algorithm improved sensitivity for confirming MBL production to 100%. For the remaining C-T positive but I-R negative isolates, C-E showed 75% sensitivity and 78.6% specificity in detecting ESBL production. The DDST further confirmed ESBL production in six out of nine ESBL producers (66.6%). In conclusion, we established a simple and cost-effective diagnostic algorithm, enabling screening and confirmation of carbapenemase and ESBL production in CRPA.IMPORTANCECarbapenem-resistant Pseudomonas aeruginosa (CRPA) is a major global health threat, and rapid identification of its resistance mechanisms is crucial for effective treatment and infection control. Differentiating between carbapenemase-producing (CP), extended-spectrum β-lactamase-producing (ESBL), and AmpC-hyperproducing CRPA is challenging, as conventional susceptibility testing cannot reliably distinguish these resistance mechanisms. Our study presents a simple, cost-effective, and easy-to-implement phenotypic diagnostic algorithm that enables accurate screening and confirmation of CP and ESBL production in CRPA. This method is particularly valuable for laboratories lacking access to molecular diagnostics, as it provides a practical alternative for routine testing. By facilitating the early detection of resistant P. aeruginosa strains, this approach has the potential to improve patient outcomes, optimize antimicrobial therapy, and enhance global surveillance efforts against multidrug-resistant pathogens.https://journals.asm.org/doi/10.1128/spectrum.03196-24CRPAcarbapenemase productioncarbapenemase detectionESBL detection |
| spellingShingle | Stefano Mancini Laia Garcia-Verellen Helena M. B. Seth-Smith Peter M. Keller Natalia Kolesnik-Goldmann Muhammad Ali Syed Irfan Ullah Vladimira Hinic Tim Roloff Adrian Egli Oliver Nolte Diagnostic algorithm for the detection of carbapenemases and extended-spectrum β-lactamases in carbapenem-resistant Pseudomonas aeruginosa Microbiology Spectrum CRPA carbapenemase production carbapenemase detection ESBL detection |
| title | Diagnostic algorithm for the detection of carbapenemases and extended-spectrum β-lactamases in carbapenem-resistant Pseudomonas aeruginosa |
| title_full | Diagnostic algorithm for the detection of carbapenemases and extended-spectrum β-lactamases in carbapenem-resistant Pseudomonas aeruginosa |
| title_fullStr | Diagnostic algorithm for the detection of carbapenemases and extended-spectrum β-lactamases in carbapenem-resistant Pseudomonas aeruginosa |
| title_full_unstemmed | Diagnostic algorithm for the detection of carbapenemases and extended-spectrum β-lactamases in carbapenem-resistant Pseudomonas aeruginosa |
| title_short | Diagnostic algorithm for the detection of carbapenemases and extended-spectrum β-lactamases in carbapenem-resistant Pseudomonas aeruginosa |
| title_sort | diagnostic algorithm for the detection of carbapenemases and extended spectrum β lactamases in carbapenem resistant pseudomonas aeruginosa |
| topic | CRPA carbapenemase production carbapenemase detection ESBL detection |
| url | https://journals.asm.org/doi/10.1128/spectrum.03196-24 |
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