title: DNA methylase modifications and other linezolid resistance mutations in coagulase-negative staphylococci in Italy.
abstract: BACKGROUND
  Despite 10 years of clinical use, linezolid resistance in Staphylococcus aureus and coagulase-negative staphylococci (CoNS) is still a rare phenomenon. This study reports the mechanisms of resistance and strain types seen in clusters of linezolid-resistant CoNS from two different hospitals in Italy during the period 2008-09.
  
  
  METHODS
  Genes associated with linezolid resistance were subjected to molecular analysis and isolates were characterized by PFGE macrorestriction analysis using SmaI.
  
  
  RESULTS
  Thirty-three linezolid-resistant isolates of methicillin-resistant CoNS comprising Staphylococcus epidermidis (24), Staphylococcus hominis (5) and Staphylococcus simulans (4) were studied. The isolates showed varying levels of linezolid resistance. Almost all isolates for which linezolid MICs were 64 mg/L possessed point mutations in domain V of 23S rRNA, while isolates for which the MICs were 256 mg/L expressed methylase activity at position A2503 mediated by the cfr gene. Overall, the isolates showed reduced susceptibility to vancomycin (MICs 1-2 mg/L) and 11 of the 33 isolates showed no susceptibility to teicoplanin. These strains were also resistant to chloramphenicol (28 of 33), lincomycin (24 of 33), erythromycin (17 of 33) and quinupristin/dalfopristin (13 of 33). S. epidermidis isolates, showing mutations or methylase modifications, belonged to different PFGE profiles and to two different sequence types (ST2 and ST23), in which the cfr gene was carried on a plasmid of ∼50 kb.
  
  
  CONCLUSIONS
  Clinical CoNS strains with resistance to linezolid and other second-line antibiotics, as well as reduced susceptibility to glycopeptides, have emerged in Italy.
authors.affiliation: Department of Microbiology, University of Catania, Via Androne 81, 95124 Catania, Italy.
authors.forename: Dafne
authors.initials: D
authors.lastname: Bongiorno
authors.forename: Floriana
authors.initials: F
authors.lastname: Campanile
authors.forename: Gino
authors.initials: G
authors.lastname: Mongelli
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Baldi
authors.forename: Rosamaria
authors.initials: R
authors.lastname: Provenzani
authors.forename: Silvia
authors.initials: S
authors.lastname: Reali
authors.forename: Carolina
authors.initials: C
authors.lastname: Lo Russo
authors.forename: Maria
authors.initials: M
authors.lastname: Santagati
authors.forename: Stefania
authors.initials: S
authors.lastname: Stefani
mesh_terms: D000081:Acetamides / Q000494:pharmacology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D015373:Bacterial Typing Techniques; D016172:DNA Fingerprinting; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D024901:Drug Resistance, Multiple, Bacterial; D016521:Electrophoresis, Gel, Pulsed-Field; D006761:Hospitals; D006801:Humans; D007558:Italy; D000069349:Linezolid; D008826:Microbial Sensitivity Tests; D023303:Oxazolidinones / Q000494:pharmacology*; D017354:Point Mutation; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D013203:Staphylococcal Infections / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000145:classification / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D013212:Staphylococcus epidermidis / Q000145:classification / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D041181:Staphylococcus hominis / Q000145:classification / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics
chemical_list: D000081:Acetamides; D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D023303:Oxazolidinones; D012338:RNA, Ribosomal, 23S; D015254:DNA Modification Methylases; C025651:CCCGGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D000069349:Linezolid