The aim of the present study is to describe the ongoing spread of the KPC-producing strains, which is evolving to an epidemic in Czech hospitals. During the period of 2018-2019, a total of 108 KPC-producing Enterobacterales were recovered from 20 hospitals. Analysis of long-read sequencing data revealed the presence of several types of blaKPC-carrying plasmids; 19 out of 25 blaKPC-carrying plasmids could be assigned to R (n = 12), N (n = 5), C (n = 1) and P6 (n = 1) incompatibility (Inc) groups. Five of the remaining blaKPC-carrying plasmids were multireplicon, while one plasmid couldn't be typed. Additionally, phylogenetic analysis confirmed the spread of blaKPC-carrying plasmids among different clones of diverse Enterobacterales species. Our findings demonstrated that the increased prevalence of KPC-producing isolates was due to plasmids spreading among different species. In some districts, the local dissemination of IncR and IncN plasmids was observed. Additionally, the ongoing evolution of blaKPC-carrying plasmids, through genetic rearrangements, favours the preservation and further dissemination of these mobile genetic elements. Therefore, the situation should be monitored, and immediate infection control should be implemented in hospitals reporting KPC-producing strains.

• MeSH
• Anti-Bacterial Agents pharmacology   MeSH
• Drug Resistance, Bacterial * MeSH
• Bacterial Proteins genetics   metabolism   MeSH
• beta-Lactamases metabolism   MeSH
• Epidemics MeSH
• Klebsiella Infections epidemiology   microbiology   MeSH
• Klebsiella pneumoniae isolation & purification   metabolism   MeSH
• Humans MeSH
• Hospitals statistics & numerical data   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic epidemiology   MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Bacterial Proteins MeSH
• beta-Lactamases MeSH
• carbapenemase MeSH Browser

Ten Enterobacteriaceae isolates collected in a Czech hospital carried blaKPC-positive plasmids of different sizes (∼30, ∼45, and ∼80 kb). Sequencing revealed three types of plasmids (A to C) with the Tn4401a transposon. Type A plasmids comprised an IncR backbone and a KPC-2-encoding multidrug resistance (MDR) region. Type B plasmids were derivatives of type A plasmids carrying an IncN3-like segment, while type C plasmids were IncP6 plasmids sharing the same KPC-2-encoding MDR region with type A and B plasmids.

• Keywords
• Citrobacter freundii, Illumina sequencing, IncR, ST18, Tn4401a,
• MeSH
• Anti-Bacterial Agents therapeutic use   MeSH
• beta-Lactamases genetics   metabolism   MeSH
• Citrobacter freundii drug effects   enzymology   genetics   isolation & purification   MeSH
• Enterobacteriaceae Infections drug therapy   epidemiology   microbiology   MeSH
• Escherichia coli drug effects   enzymology   genetics   isolation & purification   MeSH
• Gene Expression MeSH
• Isoenzymes genetics   metabolism   MeSH
• Carbapenems therapeutic use   MeSH
• Klebsiella pneumoniae drug effects   enzymology   genetics   isolation & purification   MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Drug Resistance, Multiple, Bacterial genetics   MeSH
• Morganella morganii drug effects   enzymology   genetics   isolation & purification   MeSH
• Hospitals MeSH
• Open Reading Frames MeSH
• Plasmids chemistry   classification   metabolism   MeSH
• Base Sequence MeSH
• Sequence Analysis, DNA MeSH
• DNA Transposable Elements MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic epidemiology   MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• beta-lactamase KPC-2 MeSH Browser
• beta-Lactamases MeSH
• Isoenzymes MeSH
• Carbapenems MeSH
• DNA Transposable Elements MeSH

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 = 8). The E. coli isolates were of different STs, while the E. cloacae isolate belonged to ST109. Twenty-four isolates carried blaOXA-48, while two isolates carried blaOXA-181 or blaOXA-232 Almost all isolates (n = 22) carried blaOXA-48-positive plasmids of a similar size (∼60 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 blaOXA-48 Sequencing showed that the majority of blaOXA-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 blaOXA-181-carrying plasmid was identical to the IncX3 plasmid pOXA181_EC14828, while the blaOXA-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 blaOXA-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 blaOXA-48 in the Czech Republic. In combination with the difficulties with their detection, OXA-48 producers constitute an important public threat.

• Keywords
• ColE2-like, IncL, IncX3, Klebsiella pneumoniae, OXA-181, OXA-232, Tn1999.2, Tn1999.5,
• MeSH
• Anti-Bacterial Agents pharmacology   MeSH
• Chromosomes, Bacterial genetics   MeSH
• Bacterial Proteins genetics   metabolism   MeSH
• beta-Lactamases genetics   metabolism   MeSH
• Enterobacteriaceae drug effects   enzymology   genetics   MeSH
• Escherichia coli drug effects   enzymology   genetics   MeSH
• Klebsiella pneumoniae drug effects   enzymology   genetics   MeSH
• Microbial Sensitivity Tests MeSH
• Plasmids genetics   MeSH
• Gene Transfer, Horizontal genetics   MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Bacterial Proteins MeSH
• beta-Lactamases MeSH
• carbapenemase MeSH Browser