Development of a Multiplex PCR Assay for the Detection of Metallo-Beta-Lactamase Genes in Pseudomonas aeruginosa

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Alexis Luengo; Laura Navarro-Heredia; Claudia Rabert; Piedad Cortés-Cortés; Eduardo Karahanian & Gino Corsini


The appearance of Pseudomonas aeruginosa strains with multi-resistance to antibiotics is a clinical problem of great relevance. The methods for detecting these resistances are laborious and slow, which is a complication when treating patients promptly. In this work, we developed a simple method for simultaneous detection of several carbapenem resistance genes using a multiplex PCR assay. The PCR assay developed, followed by electrophoretic separation of fragments, allows to simultaneously identify the presence of 6 antibiotic resistance genes: bla-VIM (261 bp), bla-IMP (587 bp), bla-SPM (648 bp), bla-GIM-1 (753 bp), bla-NDM-1 (813 bp) and bla-KPC (882 bp). We analyzed 7 clinical isolates of P. aeruginosa obtained in Chile, finding the resistance genes bla-VIM, bla-IMP, bla-SPM, bla-GIM, and bla-NDM in 5 of them. We found a perfect correlation between the detection of various resistance genes by PCR and the results obtained by antibiograms. Interestingly, 2 of the strains possessed 3 different resistance genes simultaneously. Finally, in this work, we found the presence of 3 genes never described before in clinical isolates of P. aeruginosa in Chile (bla-IMP, bla-SPM, and bla-GIM-1). We developed a rapid multiplex PCR test for the simultaneous detection of up to 6 antibiotic resistance genes of the metallo-β-lactamase family in P. aeruginosa.

KEY WORDS: Antibiotic resistance; P. aeruginosa; Multiplex PCR; Carbapenem.

How to cite this article

LUENGO, A.; NAVARRO-HEREDIA, L.; RABERT, C.; CORTÉS-CORTÉS, P.; KARAHANIAN, E. & CORSINI, G. Development of a multiplex PCR assay for the detection of metallo-beta-lactamase genes in Pseudomonas aeruginosa. Int. J. Morphol., 41(2):466-470, 2023.