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Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes
Published on: March 3, 2023
Evaluation of phenotypic detection of carbapenemase-producing Pseudomonas spp. from clinical isolates
Ivson Cassiano de Oliveira Santos1, Orlando Carlos da Conceiçāo Neto1, Bianca Santos da Costa1
1Laboratório de Pesquisa Em Infecção Hospitalar (LAPIH), Instituto Oswaldo Cruz - FIOCRUZ, Fundação Oswaldo Cruz, Av. Brasil, 4365, Rio de Janeiro, RJ, 21045900, Brazil.
Abstract:
Carbapenems are considered last-resort antibiotics for the treatment of infections caused by multidrug-resistant Gram-negative bacteria. Although the main mechanism of carbapenem-resistance in Pseudomonas aeruginosa is the loss of OprD porin, carbapenemases continue to be a problem worldwide. The aim of this study was to evaluate the performance of phenotypic tests (Carba NP, Blue Carba, and mCIM/eCIM) for detection of carbapenemase-producing Pseudomonas spp. in Brazil. One hundred twenty-seven Pseudomonas spp. clinical isolates from different Brazilian states were submitted to phenotypic and molecular carbapenemase detection. A total of 90 carbapenemase-producing P. aeruginosa and 5 Pseudomonas putida (35, blaVIM-2; 17, blaSPM-1; 2, blaIMP-10; 1, blaVIM-24; 1, blaNDM-1; 39, blaKPC-2). The phenotypic Carba NP, Blue Carba, and mCIM/eCIM showed sensitivity of 94.7%, 93.6%, and 93.6%, and specificity of 90.6%, 100%, and 96.8%, respectively. However, only the Carba NP presented the highest sensitivity and showed the ability in differentiating the carbapenemases between class A and class B using EDTA. Blue Carba failed to detect most of the class B carbapenemases, having the worst performance using EDTA. Our results show changes in the epidemiology of the spread of carbapenemases and the importance of their detection by phenotypic and genotypic tests. Such, it is essential to use analytical tools that faithfully detect bacterial resistance in vitro in a simple, sensitive, rapid, and cost-effective way. Much effort must be done to improve the current tests and for the development of new ones.
Insights
Phenotypic tests like Carba NP effectively detect carbapenemase-producing Pseudomonas in Brazil. Carba NP showed high sensitivity and specificity, differentiating carbapenemase classes, unlike Blue Carba.
Area of Science:
- Clinical Microbiology
- Antimicrobial Resistance
- Molecular Biology
Background:
- Carbapenems are crucial last-resort antibiotics for multidrug-resistant Gram-negative infections.
- Carbapenemases are a growing global threat, particularly in Pseudomonas aeruginosa.
- Accurate detection of carbapenemase production is vital for effective treatment strategies.
Purpose of the Study:
- To evaluate the performance of phenotypic tests (Carba NP, Blue Carba, mCIM/eCIM) for detecting carbapenemase-producing Pseudomonas spp. in Brazil.
- To compare the sensitivity and specificity of these phenotypic tests against molecular methods.
- To assess the ability of phenotypic tests to differentiate carbapenemase classes.
Main Methods:
- Collected 127 Pseudomonas spp. clinical isolates from Brazil.
- Performed phenotypic carbapenemase detection using Carba NP, Blue Carba, and mCIM/eCIM tests.
- Utilized molecular methods for carbapenemase gene detection and confirmation.
Main Results:
- Carba NP demonstrated high sensitivity (94.7%) and specificity (90.6%).
- Blue Carba and mCIM/eCIM showed high specificity (100% and 96.8%) but slightly lower sensitivity (93.6%).
- Carba NP successfully differentiated between carbapenemase classes A and B using EDTA, while Blue Carba struggled with class B enzymes.
Conclusions:
- Phenotypic tests, particularly Carba NP, are valuable tools for detecting carbapenemase-producing Pseudomonas in Brazil.
- The study highlights evolving carbapenemase epidemiology and the need for reliable detection methods.
- Continued development of simple, sensitive, rapid, and cost-effective diagnostic tests is essential.
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