Rapid identification and differentiation of blaGES cephalosporinase and carbapenemase types using real-time PCR assay
Objectives: Guiana extended-spectrum β-lactamase (blaGES) is a β-lactamase with at least 57 known variants. Based on amino acid variation at position 170, they can be classified into variants with cephalosporinase activity and variants with carbapenemase activity. This study aimed to develop a rapid...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-06-01
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| Series: | Journal of Global Antimicrobial Resistance |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213716525001018 |
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| Summary: | Objectives: Guiana extended-spectrum β-lactamase (blaGES) is a β-lactamase with at least 57 known variants. Based on amino acid variation at position 170, they can be classified into variants with cephalosporinase activity and variants with carbapenemase activity. This study aimed to develop a rapid and efficient method for the detection and identification of the different functional blaGES variants. Methods: We developed a real-time PCR assay to screen for the presence of all blaGES variants and a second allele-specific PCR targeting the amino acid at position 170 for rapid differentiation of the cephalosporinase and carbapenemase variants. Results: When applied to 100 isolates characterized before by whole genome sequencing, the blaGES screening assay demonstrated a sensitivity and specificity of 100%. The blaGES differentiation assay correctly classified the positive samples and also allowed identification of isolates containing both a cephalosporinase and a carbapenemase blaGES variant. The results were reproducible down to approximately 5 cells per PCR. Conclusions: We have developed a rapid, sensitive and easily applicable method to detect and distinguish glycine, serine and asparagine blaGES variants using real-time PCR assays. These assays are suitable for routine use on cultured isolates but could, after further validation, even be used directly on screening broths. The assays will facilitate the detection and characterization of blaGES for both diagnostic purposes, antibiotic stewardship, infection prevention and control, and research purposes. |
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| ISSN: | 2213-7165 |