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Antimicrob Agents Chemother. 2017 Aug 24;61(9). pii: e00869-17. doi: 10.1128/AAC.00869-17. Print 2017 Sep.

Tandem Amplification of the Staphylococcal Cassette Chromosome mec Element Can Drive High-Level Methicillin Resistance in Methicillin-Resistant Staphylococcus aureus.

Author information

1
Department of Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
2
School of Mathematics, Statistics and Applied Mathematics, National University of Ireland, Galway, Ireland.
3
School of Biotechnology, Dublin City University, Dublin, Ireland.
4
Roslin Institute, The University of Edinburgh, Easter Bush, Midlothian, Scotland, United Kingdom.
5
Department of Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland gerard.fleming@nuigalway.ie jamesp.ogara@nuigalway.ie.

Abstract

Hospital-associated methicillin-resistant Staphylococcus aureus (MRSA) strains typically express high-level, homogeneous (HoR) β-lactam resistance, whereas community-associated MRSA (CA-MRSA) more commonly express low-level heterogeneous (HeR) resistance. Expression of the HoR phenotype typically requires both increased expression of the mecA gene, carried on the staphylococcal cassette chromosome mec element (SCCmec), and additional mutational event(s) elsewhere on the chromosome. Here the oxacillin concentration in a chemostat culture of the CA-MRSA strain USA300 was increased from 8 μg/ml to 130 μg/ml over 13 days to isolate highly oxacillin-resistant derivatives. A stable, small-colony variant, designated HoR34, which had become established in the chemostat culture was found to have acquired mutations in gdpP, clpX, guaA, and camS Closer inspection of the genome sequence data further revealed that reads covering SCCmec were ∼10 times overrepresented compared to other parts of the chromosome. Quantitative PCR (qPCR) confirmed >10-fold-higher levels of mecA DNA on the HoR34 chromosome, and MinION genome sequencing verified the presence of 10 tandem repeats of the SCCmec element. qPCR further demonstrated that subculture of HoR34 in various concentrations of oxacillin (0 to 100 μg/ml) was accompanied by accordion-like contraction and amplification of the SCCmec element. Although slower growing than strain USA300, HoR34 outcompeted the parent strain in the presence of subinhibitory oxacillin. These data identify tandem amplification of the SCCmec element as a new mechanism of high-level methicillin resistance in MRSA, which may provide a competitive advantage for MRSA under antibiotic selection.

KEYWORDS:

SCCmec; Staphylococcus; antibiotic; antibiotic resistance; beta-lactams; continuous culture; mecA beta-lactam; mechanism; resistance

PMID:
28717043
PMCID:
PMC5571284
DOI:
10.1128/AAC.00869-17
[Indexed for MEDLINE]
Free PMC Article

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