Elizeth Lopes1, Maria José Saavedra2, Eliana Costa3, Hermínia de Lencastre4, Laurent Poirel5, Marta Aires-de-Sousa6. 1. Laboratory of Molecular Genetics, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Lisbon, Portugal. 2. Laboratory Medical Microbiology, Department of Veterinary Sciences, CITAB-Centre for the Research and Technology Agro-Environmental and Biological Sciences, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal. 3. Centro Hospitalar de Trás-os-Montes e Alto Douro, Serviço de Patologia Clínica, Vila Real, Portugal. 4. Laboratory of Molecular Genetics, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Lisbon, Portugal; Laboratory of Microbiology and Infectious Diseases, The Rockefeller University, NY, USA. 5. Medical and Molecular Microbiology Unit, Faculty of Science and Medicine, University of Fribourg, Fribourg, Switzerland. 6. Laboratory of Molecular Genetics, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Lisbon, Portugal; Escola Superior de Saúde da Cruz Vermelha Portuguesa, Lisbon, Portugal. Electronic address: msousa@esscvp.eu.
Abstract
OBJECTIVES: To provide, for the first time, data on the molecular epidemiology of carbapenemase-producing Klebsiella pneumoniae clinical isolates from the northern region of Portugal (Trás-os-Montes and Alto Douro). METHODS: A total of 106 carbapenemase-producing K. pneumoniae isolates recovered from clinical samples and rectal swabs between January 2018 and March 2019 were included in this study. All isolates were characterized by antimicrobial susceptibility, identification of resistance determinants, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and plasmid analysis. RESULTS: The most common carbapenemase identified was KPC-2 (91%), followed by OXA-48 (9%). The blaKPC-2 gene was carried onto IncN (60%) and IncF (40%) plasmid types, whereas the blaOXA-48 gene was mainly located on the IncL (90%) incompatibility group. Molecular characterization distributed the 106 isolates into 29 PFGE types and 21 sequence types (STs), but three clones included 50% of the isolates: PFGE A-ST147-KPC-2 (29%), B-ST15-KPC-2 (15%), and C-ST11-OXA-48 (6%). Antimicrobial resistance rates were the following: ciprofloxacin (76%), trimethoprim-sulfamethoxazole (75%), tobramycin (62%), gentamicin (34%), amikacin (25%), tigecycline (21%), fosfomycin (10%), and colistin (7%). None of the colistin-resistant isolates harboured mcr genes. All isolates remained susceptible to ceftazidime/avibactam, but 10% presented elevated MICs (3 and 4mg/L). CONCLUSIONS: KPC-2 was the predominant carbapenemase among K. pneumoniae isolates currently circulating at this hospital from northern Portugal, followed by OXA-48. These data contrast with those obtained from the rest of the country, where KPC-3 predominates. This study showed a polyclonal structure of KPC-2-producing K. pneumoniae isolates with a predominance of the ST147 and ST15 clones.
OBJECTIVES: To provide, for the first time, data on the molecular epidemiology of carbapenemase-producing Klebsiella pneumoniae clinical isolates from the northern region of Portugal (Trás-os-Montes and Alto Douro). METHODS: A total of 106 carbapenemase-producing K. pneumoniae isolates recovered from clinical samples and rectal swabs between January 2018 and March 2019 were included in this study. All isolates were characterized by antimicrobial susceptibility, identification of resistance determinants, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and plasmid analysis. RESULTS: The most common carbapenemase identified was KPC-2 (91%), followed by OXA-48 (9%). The blaKPC-2 gene was carried onto IncN (60%) and IncF (40%) plasmid types, whereas the blaOXA-48 gene was mainly located on the IncL (90%) incompatibility group. Molecular characterization distributed the 106 isolates into 29 PFGE types and 21 sequence types (STs), but three clones included 50% of the isolates: PFGE A-ST147-KPC-2 (29%), B-ST15-KPC-2 (15%), and C-ST11-OXA-48 (6%). Antimicrobial resistance rates were the following: ciprofloxacin (76%), trimethoprim-sulfamethoxazole (75%), tobramycin (62%), gentamicin (34%), amikacin (25%), tigecycline (21%), fosfomycin (10%), and colistin (7%). None of the colistin-resistant isolates harboured mcr genes. All isolates remained susceptible to ceftazidime/avibactam, but 10% presented elevated MICs (3 and 4mg/L). CONCLUSIONS:KPC-2 was the predominant carbapenemase among K. pneumoniae isolates currently circulating at this hospital from northern Portugal, followed by OXA-48. These data contrast with those obtained from the rest of the country, where KPC-3 predominates. This study showed a polyclonal structure of KPC-2-producing K. pneumoniae isolates with a predominance of the ST147 and ST15 clones.
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