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Our study sought to perform a genome-wide analysis of multiclonal \n \n K. pneumoniae<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> isolates (n<\/jats:italic>=39; 23 patients) producing extended spectrum beta-lactamases and\/or carbapenemases sourced between 2011 and 2016 in a Portuguese tertiary-care hospital. All isolates showed resistance to third-generation cephalosporins and six isolates (five patients) were also carbapenem resistant. Genome-wide-based phylogenetic analysis revealed a topology representing ongoing dissemination of three main sequence-type (ST) clades (ST15, ST147 and ST307) and transmission across different wards, compatible with missing links that can take the form of undetected colonized patients. Two carbapenemase-coding genes were detected: blaKPC-3<\/jats:sub>\n <\/jats:italic>, located on a Tn4401d<\/jats:italic> transposon, and blaGES-5<\/jats:sub>\n <\/jats:italic> on a novel class 3 integron. Additionally, four genes coding for ESBLs (blaBEL-1<\/jats:sub>\n <\/jats:italic>, blaCTX-M-8<\/jats:sub>\n <\/jats:italic>, blaCTX-M-15<\/jats:sub>\n <\/jats:italic> and blaCTX-M-32<\/jats:sub>\n <\/jats:italic>) were also detected. ESBL horizontal dissemination across five clades is highlighted by the similar genetic environments of blaCTX-M-15<\/jats:sub>\n <\/jats:italic> gene upstream of ISEcp1<\/jats:italic> on a Tn3<\/jats:italic>-like transposon. Overall, this study provides a high-resolution genome-wide perspective on the epidemiology of ESBL and carbapenemase-producing \n \n K. pneumoniae<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> in a healthcare setting while contributing for the adoption of appropriate intervention and prevention strategies.<\/jats:p>","DOI":"10.1099\/mgen.0.000349","type":"journal-article","created":{"date-parts":[[2020,4,1]],"date-time":"2020-04-01T16:16:42Z","timestamp":1585757802000},"source":"Crossref","is-referenced-by-count":10,"title":["Whole-genome sequencing resolves a polyclonal outbreak by extended-spectrum beta-lactam and carbapenem-resistant Klebsiella pneumoniae in a Portuguese tertiary-care hospital"],"prefix":"10.1099","volume":"7","author":[{"given":"Jo\u00e3o","family":"Perdig\u00e3o","sequence":"first","affiliation":[{"name":"Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Portugal"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2579-1799","authenticated-orcid":false,"given":"Ana","family":"Modesto","sequence":"additional","affiliation":[{"name":"Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Portugal"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A. 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