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* Department of Biology and Biochemistry, University of Bath, Bath
BA2 7AY, United Kingdom; Edited by Christopher T. Walsh, Harvard Medical School, Boston,
MA, and approved April 16, 2002 (received for review February 22, 2002)
Methicillin-resistant Staphylococcus aureus (MRSA)
is a major cause of hospital-acquired infections that are becoming
increasingly difficult to combat because of emerging resistance to all
current antibiotic classes. The evolutionary origins of MRSA are poorly understood, no rational nomenclature exists, and there is no consensus on the number of major MRSA clones or the relatedness of clones described from different countries. We resolve all of these issues and
provide a more thorough and precise analysis of the evolution of MRSA
clones than has previously been possible. Using multilocus sequence
typing and an algorithm, BURST, we analyzed an
international collection of 912 MRSA and methicillin-susceptible
S. aureus (MSSA) isolates. We identified 11 major MRSA
clones within five groups of related genotypes. The putative ancestral
genotype of each group and the most parsimonious patterns of descent of
isolates from each ancestor were inferred by using BURST,
which, together with analysis of the methicillin resistance genes,
established the likely evolutionary origins of each major MRSA clone,
the genotype of the original MRSA clone and its MSSA progenitor, and the extent of acquisition and horizontal movement of the methicillin resistance genes. Major MRSA clones have arisen repeatedly from successful epidemic MSSA strains, and isolates with decreased susceptibility to vancomycin, the antibiotic of last resort, are arising from some of these major MRSA clones, highlighting a depressing progression of increasing drug resistance within a small number of
ecologically successful S. aureus genotypes.
Microbiology
The evolutionary history of methicillin-resistant
Staphylococcus aureus (MRSA)
,
, Institute of Molecular
Medicine, John Radcliffe Hospital, Oxford, OX3 9DU, United Kingdom;
§ Division of Microbiology and Infectious Diseases,
University Hospital Nottingham, Nottingham, NG7 2UH, United
Kingdom; and ¶ Department of Infectious Disease
Epidemiology, Faculty of Medicine, Imperial College, St. Mary's
Hospital, London W2 1PG, United Kingdom
To whom reprint requests should be addressed. E-mail:
m.c.enright{at}bath.ac.uk.
www.pnas.org/cgi/doi/10.1073/pnas.122108599
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|
|
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|
|
|
|
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|
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|
|
|
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|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
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|
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