title: Molecular analysis of linezolid resistance in clinical Enterococcus faecium isolates by polymerase chain reaction and pyrosequencing.
abstract: Resistance to linezolid has been associated with a G2576T mutation in the 23 S rRNA gene. Clinical isolates of linezolid-sensitive and linezolid-resistant vancomycin-resistant Enterococcus faecium of a liver transplant patient have been analysed for the G2576T mutation by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), conventional sequencing and pyrosequencing. A clear association between the number of mutated 23 S rRNA genes and the level of linezolid resistance has been demonstrated. Linezolid susceptibility re-emerged after cessation of linezolid therapy; however, the re-initiation of linezolid therapy resulted in the re-emergence of linezolid-resistant strains. Pyrosequencing rapidly detected the number of mutated alleles and is superior to conventional PCR-RFLP for the detection of heterozygous mutations.
authors.affiliation: Department of Infectious Diseases, Virology, University of Heidelberg, Heidelberg, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Schnitzler
authors.forename: K
authors.initials: K
authors.lastname: Schulz
authors.forename: C
authors.initials: C
authors.lastname: Lampson
authors.forename: M
authors.initials: M
authors.lastname: Geiss
authors.forename: H K
authors.initials: HK
authors.lastname: Geiss
mesh_terms: D000081:Acetamides / Q000494:pharmacology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D016984:Enterococcus faecium / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D000069349:Linezolid; D023303:Oxazolidinones / Q000494:pharmacology*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D000081:Acetamides; D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D023303:Oxazolidinones; D012338:RNA, Ribosomal, 23S; D000069349:Linezolid
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