AUTHOR=Marí-Almirall Marta , Ferrando Núria , Fernández Mariana José , Cosgaya Clara , Viñes Joaquim , Rubio Elisa , Cuscó Anna , Muñoz Laura , Pellice Martina , Vergara Andrea , Campo Irene , Rodríguez-Serna Laura , Santana Gemina , Del Río Ana , Francino Olga , Ciruela Pilar , Ballester Frederic , Marco Francesc , Martínez José Antonio , Soriano Álex , Pitart Cristina , Vila Jordi , Roca Ignasi ,  MERCyCAT Study Group , Jove Pepa Pérez , Padilla Emma , Ballestero-Téllez Mónica , Zboromyrska Yuliya , Benítez Miguel Ángel , Clivillé Raquel , González Sabina , Calvet Iolanda , Gallés Carmen , Sauca Goretti , Martí-Sala Carmina , Pulido Mᵃ Angeles , Vilamala Anna , González-Cuevas Araceli , Gené Amadeu , Trujillo Gloria , Madueño Joan Lopez , Raga Xavier , Ester Picó Frederic Gómez , Sarvisé Carolina , Pujol Isabel , Font Xesca 

TITLE=Clonal Spread and Intra- and Inter-Species Plasmid Dissemination Associated With Klebsiella pneumoniae Carbapenemase-Producing Enterobacterales During a Hospital Outbreak in Barcelona, Spain

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.781127

DOI=10.3389/fmicb.2021.781127

ISSN=1664-302X

ABSTRACT=<p><bold>Objectives:</bold> The study aimed to characterize the clonal spread of resistant bacteria and dissemination of resistance plasmids among carbapenem-resistant Enterobacterales at a tertiary hospital in Catalonia, Spain.</p><p><bold>Methods:</bold> Isolates were recovered from surveillance rectal swabs and diagnostic samples. Species identification was by matrix-assisted laser desorption ionization-time time of flight mass spectrometry (MALDI-TOF MS). Molecular typing was performed by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Antimicrobial susceptibility was assessed by gradient-diffusion and carriage of <italic>bla</italic> genes was detected by PCR. Plasmid typing, conjugation assays, S1-PFGE studies and long-read sequencing were used to characterize resistance plasmids.</p><p><bold>Results:</bold> From July 2018 to February 2019, 125 <italic>Klebsiella pneumoniae</italic> carbapenemase (KPC)-producing Enterobacterales were recovered from 101 inpatients from surveillance (74.4%) or clinical samples (25.6%), in a tertiary hospital in Barcelona. Clonality studies identified a major clone of <italic>Klebsiella pneumoniae</italic> belonging to sequence type ST15 and additional isolates of <italic>K. pneumoniae</italic>, <italic>Escherichia coli</italic> and <italic>Enterobacter</italic> sp. from different STs. All isolates but one carried the <italic>bla</italic><sub>KPC–2</sub> allelic variant. The <italic>bla</italic><sub>KPC–2</sub> gene was located in an IncFIIk plasmid of circa 106 Kb in a non-classical Tn<italic>4401</italic> element designated NTE<sub>KPC</sub>-pMC-2-1. Whole-genome sequencing revealed different rearrangements of the 106 Kb plasmid while the NTE<sub>KPC</sub>-pMC-2-1 module was highly conserved.</p><p><bold>Conclusion:</bold> We report a hospital outbreak caused by the clonal dissemination of KPC-producing ST15 <italic>K. pneumoniae</italic> but also the intra- and inter-species transmission of the <italic>bla</italic><sub>KPC–2</sub> gene associated with plasmid conjugation and/or transposon dissemination. To our knowledge, this is the first report of an outbreak caused by KPC-producing Enterobacterales isolated from human patients in Catalonia and highlights the relevance of surveillance studies in the early detection and control of antibiotic resistant high-risk clones.</p>