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The <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> gene is frequently embedded in Tn4401 transposon, and less frequently in non-Tn4401 elements (NTE<jats:sub>KPC<\/jats:sub>) variants I-III. The first case of KPC in the UC-CHRISTUS Clinical Hospital was detected in <jats:italic>Pseudomonas aeruginosa<\/jats:italic>. Soon after this event, KPC was detected in 2 additional <jats:italic>Pseudomonas aeruginosa<\/jats:italic>, 3 <jats:italic>Escherichia coli<\/jats:italic>, 3 <jats:italic>Enterobacter cloacae<\/jats:italic>, 3 <jats:italic>Klebsiella pneumoniae,<\/jats:italic> and 1 <jats:italic>Citrobacter freundii<\/jats:italic>, isolated from 6 different patients. We aimed to elucidate the possible mechanisms of genetic transfer and dissemination of the <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> gene among isolates of this multispecies outbreak. A molecular epidemiology analysis of the above mentioned clinical isolates (<jats:italic>n<\/jats:italic>\u2009=\u200913) through Multi-Locus Sequence Typing, plasmid analysis, Pulsed-Field Gel-Electrophoresis, and Whole-genome sequencing (WGS) was performed.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>High-risk sequence types were found: <jats:italic>K. pneumoniae<\/jats:italic> ST11, <jats:italic>P. aeruginosa<\/jats:italic> ST654, and <jats:italic>E. cloacae<\/jats:italic> ST114. All enterobacterial isolates were not clonal except for 3 <jats:italic>E. coli<\/jats:italic> isolated from the same patient. WGS analysis in 6 enterobacterial isolates showed that 4 of them had <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> embedded in a novel variant of NTE<jats:sub>KPC<\/jats:sub> designated NTE<jats:sub>KPC<\/jats:sub>-IIe. Upstream of <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> gene there was a 570 pb truncated <jats:italic>bla<\/jats:italic><jats:sub>TEM-1<\/jats:sub> gene followed by an insertion sequence that was 84% similar to ISEc63, a 4473\u2009bp element of the Tn3 family. Downstream the <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> gene there was a truncated ISKpn6 gene, and the inverted repeat right sequence of Tn4401. The ISec63-like element together with the <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> gene plus Tn4401 remnants were inserted in the Tra operon involved in conjugative transfer of the plasmid. This NTE was carried in a broad host-range IncN plasmid. <jats:italic>P. aeruginosa<\/jats:italic> isolates carried <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> gene embedded in a typical Tn4401b transposon in a different plasmid, suggesting that there was no plasmid transfer between <jats:italic>Enterobacteriaceae<\/jats:italic> and <jats:italic>P. aeruginosa<\/jats:italic> as initially hypothesized.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>Most enterobacterial isolates had <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> embedded in the same NTE<jats:sub>KPC<\/jats:sub>-IIe element, suggesting that this multispecies KPC outbreak was due to horizontal gene transfer rather than clonal spread. This poses a greater challenge to infection control measures often directed against containment of clonal spread.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s12866-021-02169-3","type":"journal-article","created":{"date-parts":[[2021,4,9]],"date-time":"2021-04-09T11:03:31Z","timestamp":1617966211000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["A multispecies outbreak of carbapenem-resistant bacteria harboring the blaKPC gene in a non-classical transposon element"],"prefix":"10.1186","volume":"21","author":[{"given":"Aniela","family":"Wozniak","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristian","family":"Figueroa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francisco","family":"Moya-Flores","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Piero","family":"Guggiana","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Castillo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lina","family":"Rivas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 M.","family":"Munita","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia C.","family":"Garc\u00eda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,4,9]]},"reference":[{"issue":"8","key":"2169_CR1","doi-asserted-by":"publisher","first-page":"1290","DOI":"10.1093\/cid\/cix893","volume":"66","author":"RA Bonomo","year":"2018","unstructured":"Bonomo RA, Burd EM, Conly J, Limbago BM, Poirel L, Segre JA, et al. 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