Journal
MICROBIAL DRUG RESISTANCE
Volume 24, Issue 7, Pages 973-979Publisher
MARY ANN LIEBERT, INC
DOI: 10.1089/mdr.2017.0168
Keywords
OXA-48; Klebsiella pneumoniae; Croatia; IncL; M; Tn1999; 2
Funding
- Interdisciplinary Section for Antibiotic Resistance Control (ISKRA) of the Croatian Ministry of Health
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Here, we report a retrospective study conducted to elucidate emergence, epidemiology, and molecular mechanisms of resistance underlying the early spread of OXA-48 carbapenemase-producing Enterobacteriaceae in Croatia. Retrospective screening for OXA-48 producers was performed on a collection of 296 nonrepetitive, carbapenem-nonsusceptible enterobacterial isolates collected from January 2011 to December 2012 from 40 participating centers in Croatia. Antimicrobial susceptibility profiles and production of carbapenemases were assessed phenotypically. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing were used for epidemiological analysis. Resistance genes were characterized by polymerase chain reaction (PCR) and sequencing. Plasmid localization of bla(OXA-48) in isolates and transconjugants was investigated by S1-PFGE and Southern hybridization. PCR mapping was used for identification of genetic platform surrounding bla(OXA-48). Out of 296 carbapenem-nonsusceptible isolates, bla(OXA-48) gene was detected in 12 Klebsiella pneumoniae isolates. All OXA-48-producing isolates showed varying resistance to carbapenems and 11 were multidrug resistant. All coproduced additional beta-lactamases, including CTX-M-15, which was detected in eight isolates. Isolates were delineated in five clonal types by PFGE corresponding to five sequence types (STs) assigned ST15, ST16, ST37, ST528, and ST1418. All OXA-48 isolates conjugated successfully and other resistance determinants were not cotransferred. bla(OXA-48) was carried on a approximate to 60kb IncL/M plasmid and was detected within Tn1999.2 composite transposon. OXA-48, a class D carbapenemase, is emerging as a potentially significant contributor among carbapenem-resistant Enterobacteriaceae in Croatia, alongside class A and B carbapenemases. Polyclonal genetic background of K. pneumoniae isolates carrying approximate to 60kb incL/M plasmid indicates that dissemination of the bla(OXA48) gene is not driven exclusively by the spread of a single clone.
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