Hospital outbreak of vancomycin-resistant enterococci caused by a single clone of Enterococcus raffinosus and several clones of Enterococcus faecium

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Hospital outbreak of vancomycin-resistant enterococci caused by a single clone of Enterococcus raffinosus and several clones of Enterococcus faecium M. Kawalec, J. Kedzierska, A. Gajda, E. Sadowy, J. Wegrzyn, S. Naser, A.B. Skotnicki, M. Gniadkowski, W. Hryniewicz Clinical Microbiology and Infection Volume 13, Issue 9, Pages 893-901 (September 2007) DOI: 10.1111/j.1469-0691.2007.01774.x Copyright © 2007 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 1 SmaI pulsed-field gel electrophoresis patterns of selected Enterococcus raffinosus isolates. Lanes: M, λ DNA Ladder (New England Biolabs, Beverly, MA, USA); 1, isolate 2229/05 (PFGE subtype a2); 2, isolate 2225/05 (a3); 3, isolate 2226/05 (a1); 4, isolate 2223/05 (a1); 5, isolate 2209/05 (a1); 6, isolate 2215/05 (a1); 7, isolate 2217/05 (a6); 8, isolate 2205/05 (a4); 9, isolate 2229/05 (a5); 10, isolate 2220/05 (a6); 11, isolate 2208/05 (a1); and 12, Enterococcus avium ATCC 14025. Clinical Microbiology and Infection 2007 13, 893-901DOI: (10.1111/j.1469-0691.2007.01774.x) Copyright © 2007 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 2 (a) Schematic representation of the structure of transposon Tn1546 from Enterococcus faecium BM4147 [18]. (b) The long-PCR (L-PCR) amplicon of Tn1546, obtained as proposed by Palepou et al. [23], used in the restriction fragment length polymorphism (RFLP) analysis with the Bsu15I (ClaI) restriction enzyme and as a probe in hybridisation studies; the distribution of ClaI restriction sites is indicated by vertical strokes. (c) The L-PCR amplicon of the vanRSHAX region, obtained according to Palepou et al. [23], used in the RFLP analysis with DdeI (location of DdeI restriction sites is indicated). (d) Bsu15I (ClaI) polymorph VI of the Tn1546-like transposon and DdeI polymorph II of the vanRSHAX gene cluster. Lanes: M1, Gene Ruler 100-bp DNA Ladder Plus (Fermentas, Vilnius, Lithuania); 1, Bsu15I (ClaI) digest of Tn1546 of E. faecium BM4147 [18]; 2, Bsu15I (ClaI) polymorph VI of the Tn1546-like transposon; 3, DdeI digest of the vanRSHAX gene cluster of E. faecium BM4147 [18]; 4, DdeI polymorph II of the vanRSHAX gene cluster; and M2, Gene Ruler 100-bp DNA Ladder (Fermentas). Clinical Microbiology and Infection 2007 13, 893-901DOI: (10.1111/j.1469-0691.2007.01774.x) Copyright © 2007 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 3 EcoRI restriction pattern of the c. 50-kb plasmid carrying the Tn1546-like element, showing the hybridisation pattern with the Tn1546 probe. Lanes: M, Gene Ruler DNA Ladder Mix (Fermentas, Vilnius, Lithuania); 1, plasmid DNA isolated from a transconjugant of Enterococcus faecium 2227/05 digested with EcoRI (plasmid c. 50 kb); and 2, hybridisation of the Tn1546 probe to the fragments of the c. 50-kb plasmid shown in lane 1. Clinical Microbiology and Infection 2007 13, 893-901DOI: (10.1111/j.1469-0691.2007.01774.x) Copyright © 2007 European Society of Clinical Infectious Diseases Terms and Conditions