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The emergence of dual CP-CRE is particularly alarming, as they can potentially compromise the efficacy of newer antibiotics, further decreasing therapeutic alternatives. Herein, we report the emergence of multiple species of CP-CRE recovered from invasive infections in Chile that simultaneously harbor\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>KPC<\/jats:sub>\n              and\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>NDM<\/jats:sub>\n              and provide an in-depth genomic characterization of these worrisome pathogens. We collected carbapenem-resistant Enterobacterales (CRE) isolates from invasive infections over a 4-year period, across 11 healthcare centers in Chile. Bacterial species and the presence of carbapenemase genes were confirmed using MALDI-TOF and PCR assays, respectively. Antimicrobial susceptibility testing was conducted through disk diffusion and broth microdilution methods. Dual CP-CRE isolates were subjected to short- and long-read whole genome sequencing to perform a detailed genomic characterization of the isolates and of the mobile genetic elements harboring the enzymes. From a total of 1,335 CRE isolates, we observed an increase in the prevalence of CP-CRE, from 11% in 2019 to 38% in 2022. A total of 11 dual CP-CRE isolates were recovered, all of them harboring\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>KPC<\/jats:sub>\n              and\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>NDM<\/jats:sub>\n              . Species corresponded to\n              <jats:italic>Escherichia coli<\/jats:italic>\n              (\n              <jats:italic>n<\/jats:italic>\n              = 6),\n              <jats:italic>Klebsiella pneumoniae<\/jats:italic>\n              (\n              <jats:italic>n<\/jats:italic>\n              = 2),\n              <jats:italic>Klebsiella oxytoca<\/jats:italic>\n              (\n              <jats:italic>n<\/jats:italic>\n              = 2), and\n              <jats:italic>Citrobacter freundii<\/jats:italic>\n              (\n              <jats:italic>n<\/jats:italic>\n              = 1). Dual CP-CRE isolates exhibited resistance to all tested \u03b2-lactams except for cefiderocol. The\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>KPC<\/jats:sub>\n              and\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>NDM<\/jats:sub>\n              encoding genes were located on independent plasmids. Platforms harboring\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>KPC<\/jats:sub>\n              were diverse and included IncN, IncF, and IncFIB plasmids. In contrast,\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>NDM-7<\/jats:sub>\n              was only found on fairly conserved IncX3 plasmids. We report that a rapid increase of CP-CRE in Chile, alongside with the emergence of multiple bacterial species of CP-CRE co-harboring\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>KPC-2\/3<\/jats:sub>\n              and\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>NDM-7<\/jats:sub>\n              , underscores a critical public health challenge. Our data suggest that the dissemination of\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>NDM-7<\/jats:sub>\n              was predominantly facilitated by IncX3 plasmids, whereas the spread of\n              <jats:italic>bla<\/jats:italic>\n              <jats:sub>KPC<\/jats:sub>\n              involved multiple plasmid backbones. Active surveillance and genomic monitoring are critical to inform public policy and curtail the spread of these highly resistant pathogens.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/aac.01205-24","type":"journal-article","created":{"date-parts":[[2024,12,5]],"date-time":"2024-12-05T14:00:43Z","timestamp":1733407243000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":17,"title":["Multispecies emergence of dual\n            <i>bla<\/i>\n            <sub>KPC\/NDM<\/sub>\n            carbapenemase-producing Enterobacterales recovered from invasive infections in Chile"],"prefix":"10.1128","volume":"69","author":[{"given":"Ana M.","family":"Quesille-Villalobos","sequence":"first","affiliation":[{"name":"Genomics & Resistant Microbes group (GeRM), Instituto de Ciencias e Innovaci\u00f3n en Medicina (ICIM), Facultad de Medicina, Cl\u00ednica Alemana, Universidad del Desarrollo","place":["Santiago, Chile"]},{"name":"Multidisciplinary Initiative for Collaborative Research on Bacterial Resistance (MICROB-R)","place":["Santiago, Chile"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Camila","family":"Solar","sequence":"additional","affiliation":[{"name":"Genomics & Resistant Microbes group (GeRM), Instituto de Ciencias e Innovaci\u00f3n en Medicina (ICIM), Facultad de Medicina, Cl\u00ednica Alemana, Universidad del Desarrollo","place":["Santiago, Chile"]},{"name":"Multidisciplinary Initiative for Collaborative Research on Bacterial Resistance (MICROB-R)","place":["Santiago, Chile"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose R. W.","family":"Mart\u00ednez","sequence":"additional","affiliation":[{"name":"Genomics & Resistant Microbes group (GeRM), Instituto de Ciencias e Innovaci\u00f3n en Medicina (ICIM), Facultad de Medicina, Cl\u00ednica Alemana, Universidad del Desarrollo","place":["Santiago, Chile"]},{"name":"Multidisciplinary Initiative for Collaborative Research on Bacterial Resistance (MICROB-R)","place":["Santiago, Chile"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lina","family":"Rivas","sequence":"additional","affiliation":[{"name":"Genomics & Resistant Microbes group (GeRM), Instituto de Ciencias e Innovaci\u00f3n en Medicina (ICIM), Facultad de Medicina, Cl\u00ednica Alemana, Universidad del Desarrollo","place":["Santiago, Chile"]},{"name":"Multidisciplinary Initiative for Collaborative Research on Bacterial Resistance (MICROB-R)","place":["Santiago, 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