title: High abundance and diversity of antimicrobial resistance determinants among early vancomycin-resistant Enterococcus faecium in Poland.
abstract: The purpose of this study was to investigate the clonal structure, antimicrobial resistance phenotypes and their determinants among early vancomycin-resistant Enterococcus faecium (VREm) isolates in Poland. Two hundred and eighty-one VREm isolates collected between 1997 and 2005 were studied. VREm isolates were characterised by multilocus sequence typing (MLST). The presence of antimicrobial resistance determinants, transposon-specific genes, IS16 and esp Efm was checked by polymerase chain reaction (PCR). Ciprofloxacin and ampicillin resistance determinants were investigated by sequencing. Two hundred and twenty-two (79 %) and 59 (21 %) VREm isolates were vanA- and vanB-positive, respectively. Among 135 representative isolates, MLST yielded 33 different sequence types (STs), of which 29 were characteristic of hospital-associated E. faecium; 128 (94.8 %) and 123 (91.1 %) isolates harboured the IS16 and esp Efm genes, and all 135 isolates were resistant to ciprofloxacin and ampicillin. Resistance to tetracycline (71.1 % isolates) was mostly associated with tetM (75.0 %) and the concomitant presence of the Tn916 integrase gene. High-level resistance to streptomycin (93.3 % of isolates) and high-level resistance to gentamicin (94.1 % of isolates) were due to ant(6')-Ia and aac(6')-Ie-aph(2″) genes, respectively, the latter of which is known to be located on various Tn4001-type transposons. Fifteen combinations of mutations in the quinolone-determining regions of GyrA and ParC were identified, including changes not previously reported, such as S83F and A84P in GyrA. Twenty-three variants of the penicillin-binding protein PBP5 occurred in the studied group, and novel insertions at amino acid positions 433 and 568 were identified. This analysis revealed the predominance of hospital-associated strains of E. faecium, carrying an abundant and divergent range of resistance determinants among early VREm isolates in Poland.
authors.affiliation: Department of Molecular Microbiology, National Medicines Institute, ul. Chełmska 30/34, 00-725 Warsaw, Poland. ewasadowy@cls.edu.pl
authors.forename: E
authors.initials: E
authors.lastname: Sadowy
authors.forename: A
authors.initials: A
authors.lastname: Sieńko
authors.forename: I
authors.initials: I
authors.lastname: Gawryszewska
authors.forename: A
authors.initials: A
authors.lastname: Bojarska
authors.forename: K
authors.initials: K
authors.lastname: Malinowska
authors.forename: W
authors.initials: W
authors.lastname: Hryniewicz
mesh_terms: D000668:Ampicillin Resistance / Q000235:genetics; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002939:Ciprofloxacin / Q000494:pharmacology; D027081:DNA Gyrase / Q000235:genetics; D027101:DNA Topoisomerase IV / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D016984:Enterococcus faecium / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D005839:Gentamicins / Q000494:pharmacology; D006801:Humans; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D011044:Poland; D013307:Streptomycin / Q000494:pharmacology; D013752:Tetracycline / Q000494:pharmacology; D014640:Vancomycin / Q000494:pharmacology; D020713:Vancomycin Resistance / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D004251:DNA Transposable Elements; D005839:Gentamicins; D002939:Ciprofloxacin; D014640:Vancomycin; D027101:DNA Topoisomerase IV; D027081:DNA Gyrase; D013752:Tetracycline; D013307:Streptomycin
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