Molecular Characterization of OXA-48-Like-Producing Enterobacteriaceae in the Czech Republic and Evidence for Horizontal Transfer of pOXA-48-Like Plasmids

Abstract

The aim of this study was to characterize the first cases and outbreaks of OXA-48-like-producing Enterobacteriaceae recovered from hospital settings in the Czech Republic. From 2013 to 2015, 22 Klebsiella pneumoniae isolates, 3 Escherichia coli isolates, and 1 Enterobacter cloacae isolate producing OXA-48-like carbapenemases were isolated from 20 patients. Four of the patients were colonized or infected by two or three different OXA-48-like producers. The K. pneumoniae isolates were classified into nine sequence types (STs), with ST101 being predominant (n ϭ 8). The E. coli isolates were of different STs, while the E. cloacae isolate belonged to ST109. Twenty-four isolates carried bla OXA-48 , while two isolates carried bla OXA-181 or bla . Almost all isolates (n ϭ 22) carried bla OXA-48 -positive plasmids of a similar size (ϳ60 kb), except the two isolates producing OXA-181 or OXA-232. In an ST45 K. pneumoniae isolate and an ST38 E. coli isolate, S1 nuclease profiling plus hybridization indicated a chromosomal location of bla . Sequencing showed that the majority of bla OXA-48 -carrying plasmids exhibited high degrees of identity with the pOXA-48-like plasmid pE71T. Additionally, two novel pE71T derivatives, pOXA-48_30715 and pOXA-48_30891, were observed. The bla OXA-181 -carrying plasmid was identical to the IncX3 plasmid pOXA181_EC14828, while the bla OXA-232 -carrying plasmid was a ColE2-type plasmid, being a novel derivative of pOXA-232. Finally, sequencing data showed that the ST45 K. pneumoniae and ST38 E. coli isolates harbored the IS1R-based composite transposon Tn6237 containing bla OXA-48 integrated into their chromosomes. These findings underlined that the horizontal transfer of pOXA-48-like plasmids has played a major role in the dissemination of bla in the Czech Republic. In combination with the difficulties with their detection, OXA-48 producers constitute an important public threat.

Plasmid evolution in carbapenemase‐producing Enterobacteriaceae: a review

“…OXA variants in K. pneumoniae are usually spread by the IncF and L/M replicon groups. L/M has been reported to spread OXA‐48 in different countries, including the United States, the Czech Republic, Romania, and Australia (Table ) . OXA‐181 has been reported in Angola and Australia on IncF, A/C, and IncX plasmid replicon groups (Fig.…”

“…The IncL/IncM incompatibility group has been associated with multiple ESBLs, AmpCs, and carbapenemases, specifically class B and D genes . Several IncL plasmids in bla NDM‐1 ‐ and bla OXA‐48 ‐expressing clinical isolates have been widely reported, some of which include pNDM‐OM (87.1 kb), pNDM‐HK (88.8 kb), E71T (63.5 kb), and pOXA‐48‐4963 (63.5 kb) . Although L/M plasmids usually harbor bla NDM and bla OXA‐48 , they are also reportedly associated with bla IMP in Enterobacteriaceae (Table and Fig.…”

Complete Nucleotide Sequences of Two VIM-1-Encoding Plasmids from Klebsiella pneumoniae and Leclercia adecarboxylata Isolates of Czech Origin

“…Plasmids were sequenced using the Illumina MiSeq platform (Illumina Inc., San Diego, CA, USA). Assembling the reads, filling sequence gaps, and analysis and annotation of the plasmid sequences were performed as described previously (9).…”

Hospital outbreak of carbapenem-resistant Enterobacteriales associated with an OXA-48 plasmid carried mostly byEscherichia coliST399

“…These antibiotics are widely used in clinical practice as they are active against extended-spectrum β-lactamase ESBL-producing Enterobacteriaceae which are resistant to other beta-lactams (Potron et al , 2011;Nordmann, Dortet and Poirel, 2012) . Acquisition by bacteria that also produce ESBL may render them resistant to all beta-lactams currently used for treatment (Nordmann, Dortet and Poirel, 2012;Poirel, Potron and Nordmann, 2012;Skalova et al , 2017) . Several classes of carbapenemase have been described, and genes are typically found on plasmids and other mobile genetic elements (MGEs) (Potron et al , 2011;Nordmann, Dortet and Poirel, 2012;Poirel, Potron and Nordmann, 2012;Findlay, Hopkins, Alvarez-Buylla, et al , 2017;Skalova et al , 2017) .…”

“…Acquisition by bacteria that also produce ESBL may render them resistant to all beta-lactams currently used for treatment (Nordmann, Dortet and Poirel, 2012;Poirel, Potron and Nordmann, 2012;Skalova et al , 2017) . Several classes of carbapenemase have been described, and genes are typically found on plasmids and other mobile genetic elements (MGEs) (Potron et al , 2011;Nordmann, Dortet and Poirel, 2012;Poirel, Potron and Nordmann, 2012;Findlay, Hopkins, Alvarez-Buylla, et al , 2017;Skalova et al , 2017) . p OXA-48 is a typical example of a plasmid carrying a gene ( bla ) conferr ing resistance to carbapenems (Poirel, Potron and Nordmann, 2012;Skalova et al , 2017) .…”