title: Investigation of Linezolid Resistance in Staphylococci and Enterococci.
abstract: The objective of this study was to investigate an apparent increase in linezolid-nonsusceptible staphylococci and enterococci following a laboratory change in antimicrobial susceptibility testing from disk diffusion to an automated susceptibility testing system. Isolates with nonsusceptible results (n = 27) from Vitek2 were subjected to a battery of confirmatory testing which included disk diffusion, Microscan broth microdilution, Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution, gradient diffusion (Etest), 23S rRNA gene sequencing, and cfr PCR. Our results show that there is poor correlation between methods and that only 70 to 75% of isolates were confirmed as linezolid resistant with alternative phenotypic testing methods (disk diffusion, Microscan broth microdilution, CLSI broth microdilution, and Etest). 23S rRNA gene sequencing identified mutations previously associated with linezolid resistance in 16 (59.3%) isolates, and the cfr gene was detected in 3 (11.1%) isolates. Mutations located at positions 2576 and 2534 of the 23S rRNA gene were most common. In addition, two previously undescribed variants (at positions 2083 and 2345 of the 23S rRNA gene) were also identified and may contribute to linezolid resistance.
authors.affiliation: Department of Pathology, Virginia Commonwealth University Medical Center, Richmond, Virginia, USA cdoern@mcvh-vcu.edu cburnham@path.wustl.edu.
authors.forename: Christopher D
authors.identifier: 0000-0003-2454-4355
authors.initials: CD
authors.lastname: Doern
authors.affiliation: Department of Pathology and the Eugene McDermott Center for Human Growth and Development, University of Texas Southwestern Medical Center, Dallas, Texas, USA Children's Medical Center, Dallas, Texas, USA.
authors.forename: Jason Y
authors.identifier: 0000-0003-3343-4780
authors.initials: JY
authors.lastname: Park
authors.affiliation: Children's Medical Center, Dallas, Texas, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Gallegos
authors.affiliation: Department of Laboratories, Barnes-Jewish Hospital, St. Louis, Missouri, USA.
authors.forename: Debbie
authors.initials: D
authors.lastname: Alspaugh
authors.affiliation: Department of Pathology and Immunology, Department of Pediatrics, and Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, USA cdoern@mcvh-vcu.edu cburnham@path.wustl.edu.
authors.forename: Carey-Ann D
authors.identifier: 0000-0002-1137-840X
authors.initials: CA
authors.lastname: Burnham
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D024881:Drug Resistance, Bacterial; D016983:Enterococcus / Q000187:drug effects* / Q000302:isolation & purification; D006801:Humans; D000069349:Linezolid / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests / Q000379:methods / Q000592:standards; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D017422:Sequence Analysis, DNA; D013210:Staphylococcus / Q000187:drug effects* / Q000302:isolation & purification
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012338:RNA, Ribosomal, 23S; D000069349:Linezolid