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J Antimicrob Chemother. 2015 Mar;70(3):658-63. doi: 10.1093/jac/dku443. Epub 2014 Nov 6.

Molecular characterization of linezolid-resistant CoNS isolates in Japan.

Author information

1
Department of Microbiology and Molecular Genetics, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan.
2
Division of Control and Treatment of Infectious Diseases, Chiba University Hospital, Chiba 260-8677, Japan.
3
Division of Laboratory Medicine, Chiba University Hospital, Chiba 260-8677, Japan.
4
Department of Bacteriology II, National Institute of Infectious Diseases, Musashimurayama, Tokyo 208-0011, Japan.
5
Department of Microbiology and Molecular Genetics, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan tomoko-y@faculty.chiba-u.jp.

Abstract

OBJECTIVES:

Linezolid has been reported to remain active against 98% of staphylococci with resistance identified in 0.05% of Staphylococcus aureus and 1.4% of CoNS. The objective of this study was to characterize the linezolid-resistance mechanisms in the linezolid-resistant CoNS strains isolated in Japan.

METHODS:

Staphylococcus capitis strains exhibiting linezolid MICs >8 mg/L isolated from inpatients between 2012 and 2014 were screened for cfr and mutations in 23S rRNA, L3 and L4 by PCR/sequencing. Isolates were also examined for mutations in the rlmN gene.

RESULTS:

S. capitis had six 23S rRNA alleles. Five S. capitis isolates displayed linezolid MICs of 8, 16 and 32 mg/L. G2576U mutations were detected in three, four or five copies of 23S rRNA in all isolates. In two isolates exhibiting the highest linezolid MIC (32 mg/L) there was a large deletion in a single copy of 23S rRNA. Repeated 10 bp sequences were found in both 16S and 23S rRNAs, suggesting deletion by recombination between the repeats. One isolate had the mutation Ala-142→Thr in the ribosomal protein L3. All linezolid-resistant isolates also demonstrated mutations in the gene encoding RlmN methyltransferase, leading to Thr-62→Met and Gly-148→Ser.

CONCLUSIONS:

Multiple mechanisms appeared to be responsible for the elevated linezolid resistance in S. capitis isolates: a G2576U mutation in different numbers of copies of 23S rRNA, loss of a single copy of 23S rRNA and a mutation in the ribosomal protein L3, suggesting the accumulation of independent mutational events.

KEYWORDS:

S. capitis; ribosomal mutations; rlmN

PMID:
25381168
DOI:
10.1093/jac/dku443
[Indexed for MEDLINE]

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