Polyphasic characterization of carbapenem-resistant Klebsiella pneumoniae clinical isolates suggests vertical transmission of the blaKPC-3 gene
Open Access
- 26 February 2021
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 16 (2), e0247058
- https://doi.org/10.1371/journal.pone.0247058
Abstract
Carbapenem-resistant Klebsiella pneumoniae are a major global threat in healthcare facilities. The propagation of carbapenem resistance determinants can occur through vertical transmission, with genetic elements being transmitted by the host bacterium, or by horizontal transmission, with the same genetic elements being transferred among distinct bacterial hosts. This work aimed to track carbapenem resistance transmission by K. pneumoniae in a healthcare facility. The study involved a polyphasic approach based on conjugation assays, resistance phenotype and genotype analyses, whole genome sequencing, and plasmid characterization by pulsed field gel electrophoresis and optical DNA mapping. Out of 40 K. pneumoniae clinical isolates recovered over two years, five were carbapenem- and multi-drug-resistant and belonged to multilocus sequence type ST147. These isolates harboured the carbapenemase encoding bla(KPC-3) gene, integrated in conjugative plasmids of 140 kbp or 55 kbp, belonging to replicon types incFIA/incFIIK or incN/incFIIK, respectively. The two distinct plasmids encoding the bla(KPC-3) gene were associated with distinct genetic lineages, as confirmed by optical DNA mapping and whole genome sequence analyses. These results suggested vertical (bacterial strain-based) transmission of the carbapenem-resistance genetic elements. Determination of the mode of transmission of antibiotic resistance in healthcare facilities, only possible based on polyphasic approaches as described here, is essential to control resistance propagation.Funding Information
- Norte Portugal Regional Operational Programme ((NORTE 2020))
- FEDER ((PTDC/CTA-AMB/28196/2017))
- National Funds from FCT – Fundação para a Ciência e a Tecnologia
- The Swedish Research Council, Sweden
- Fundação para a Ciência e a Tecnologia (FCT), Portugal
- European Regional Development Fund (ERDF) European Regional Development Fund (ERDF), through COMPETE2020 (UID/Multi/50016/2019, UID/EQU/00511/2019 and “LEPABE-2-ECO-INNOVATION” – NORTE‐01‐0145‐ FEDER‐000005)
This publication has 49 references indexed in Scilit:
- Heterogeneous staining: a tool for studies of how fluorescent dyes affect the physical properties of DNANucleic Acids Research, 2013
- Identification of acquired antimicrobial resistance genesJournal of Antimicrobial Chemotherapy, 2012
- Multilocus Sequence Typing of Total-Genome-Sequenced BacteriaJournal of Clinical Microbiology, 2012
- A single-step competitive binding assay for mapping of single DNA moleculesBiochemical and Biophysical Research Communications, 2011
- Genetic Organization of Transposase Regions Surrounding bla KPC Carbapenemase Genes on Plasmids from Klebsiella Strains Isolated in a New York City HospitalAntimicrobial Agents and Chemotherapy, 2009
- Genetic Structures at the Origin of Acquisition of the β-Lactamase bla KPC GeneAntimicrobial Agents and Chemotherapy, 2008
- Specific detection of blaVIM and blaIMP metallo-β-lactamase genes in a single real-time PCRClinical Microbiology & Infection, 2007
- Characterization of an Integron Carrying bla IMP-1 and a New Aminoglycoside Resistance Gene, aac(6′)-31 , and Its Dissemination among Genetically Unrelated Clinical Isolates in a Brazilian HospitalAntimicrobial Agents and Chemotherapy, 2007
- Antimicrobial resistance patterns in Enterobacteriaceae isolated from an urban wastewater treatment plantFEMS Microbiology Ecology, 2007
- A General Method for Detecting and Sizing Large PlasmidsAnalytical Biochemistry, 1995