Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus.

A multilocus sequence typing (MLST) scheme has been developed for Staphylococcus aureus. The sequences of internal fragments of seven housekeeping genes were obtained for 155 S. aureus isolates from patients with community-acquired and hospital-acquired invasive disease in the Oxford, United Kingdom, area. Fifty-three different allelic profiles were identified, and 17 of these were represented by at least two isolates. The MLST scheme was highly discriminatory and was validated by showing that pairs of isolates with the same allelic profile produced very similar SmaI restriction fragment patterns by pulsed-field gel electrophoresis. All 22 isolates with the most prevalent allelic profile were methicillin-resistant S. aureus (MRSA) isolates and had allelic profiles identical to that of a reference strain of the epidemic MRSA clone 16 (EMRSA-16). Four MRSA isolates that were identical in allelic profile to the other major epidemic MRSA clone prevalent in British hospitals (clone EMRSA-15) were also identified. The majority of isolates (81%) were methicillin-susceptible S. aureus (MSSA) isolates, and seven MSSA clones included five or more isolates. Three of the MSSA clones included at least five isolates from patients with community-acquired invasive disease and may represent virulent clones with an increased ability to cause disease in otherwise healthy individuals. The most prevalent MSSA clone (17 isolates) was very closely related to EMRSA-16, and the success of the latter clone at causing disease in hospitals may be due to its emergence from a virulent MSSA clone that was already a major cause of invasive disease in both the community and hospital settings. MLST provides an unambiguous method for assigning MRSA and MSSA isolates to known clones or assigning them as novel clones via the Internet.

[1]  M. Achtman,et al.  The relative contributions of recombination and mutation to the divergence of clones of Neisseria meningitidis. , 1999, Molecular biology and evolution.

[2]  Brian G. Spratt,et al.  The Three Major Spanish Clones of Penicillin-Resistant Streptococcus pneumoniae Are the Most Common Clones Recovered in Recent Cases of Meningitis in Spain , 1999, Journal of Clinical Microbiology.

[3]  B. Spratt Multilocus sequence typing: molecular typing of bacterial pathogens in an era of rapid DNA sequencing and the internet. , 1999, Current opinion in microbiology.

[4]  B. Robinson-Dunn,et al.  Emergence of Vancomycin Resistance inStaphylococcus aureus , 1999 .

[5]  B. Robinson-Dunn,et al.  Emergence of vancomycin resistance in Staphylococcus aureus. Glycopeptide-Intermediate Staphylococcus aureus Working Group. , 1999, The New England journal of medicine.

[6]  B. Spratt,et al.  Identification of Three Major Clones of Multiply Antibiotic-Resistant Streptococcus pneumoniae in Taiwanese Hospitals by Multilocus Sequence Typing , 1998, Journal of Clinical Microbiology.

[7]  B. Spratt,et al.  A multilocus sequence typing scheme for Streptococcus pneumoniae: identification of clones associated with serious invasive disease. , 1998, Microbiology.

[8]  H. de Lencastre,et al.  Intercontinental Spread of a Multidrug-Resistant MethicillinResistant Staphylococcus aureusClone , 1998, Journal of Clinical Microbiology.

[9]  W. Grubb Genetics of MRSA , 1998 .

[10]  M. Kaufmann,et al.  Assessment of Resolution and Intercenter Reproducibility of Results of Genotyping Staphylococcus aureus by Pulsed-Field Gel Electrophoresis of SmaI Macrorestriction Fragments: a Multicenter Study , 1998, Journal of Clinical Microbiology.

[11]  K. Köhrer,et al.  Typing of methicillin-resistant Staphylococcus aureus isolates from Düsseldorf by six genotypic methods. , 1998, Journal of medical microbiology.

[12]  M. Achtman,et al.  Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Wayne N. Campbell,et al.  Phenotypic and Genotypic Characterization of Nosocomial Staphylococcus aureus Isolates from Trauma Patients , 1998, Journal of Clinical Microbiology.

[14]  L. Nicolle,et al.  Rapid geographic spread of a methicillin-resistant Staphylococcus aureus strain. , 1997, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[15]  D. James,et al.  Resistance to methicillin and other antibiotics in isolates of Staphylococcus aureus from blood and cerebrospinal fluid, England and Wales, 1989–95 , 1997, The Lancet.

[16]  A. van Belkum,et al.  Simultaneous persistence of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus in a neonatal ward of a Warsaw hospital. , 1997, The Journal of hospital infection.

[17]  F. Tenover,et al.  Methicillin-resistant Staphylococcus aureus clinical strain with reduced vancomycin susceptibility. , 1997, The Journal of antimicrobial chemotherapy.

[18]  J. Steinberg,et al.  Nosocomial and community-acquired Staphylococcus aureus bacteremias from 1980 to 1993: impact of intravascular devices and methicillin resistance. , 1996, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[19]  B. Cookson,et al.  Inter-centre comparison of pulsed-field gel electrophoresis for the typing of methicillin-resistant Staphylococcus aureus. , 1996, Journal of medical microbiology.

[20]  A. Tomasz,et al.  Testing the efficacy of a molecular surveillance network: methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecium (VREF) genotypes in six hospitals in the metropolitan New York City area. The BARG Initiative Pilot Study Group. Bacterial Antibiotic Resistance , 1996, Microbial drug resistance.

[21]  D H Persing,et al.  Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing , 1995, Journal of clinical microbiology.

[22]  J. Miller,et al.  Pulsed-field gel electrophoresis as a replacement for bacteriophage typing of Staphylococcus aureus , 1995, Journal of clinical microbiology.

[23]  R. Marples,et al.  A major outbreak of methicillin-resistant Staphylococcus aureus caused by a new phage-type (EMRSA-16) , 1995, The Journal of hospital infection.

[24]  A. Tomasz,et al.  Spread and maintenance of a dominant methicillin-resistant Staphylococcus aureus (MRSA) clone during an outbreak of MRSA disease in a Spanish hospital , 1994, Journal of clinical microbiology.

[25]  Epidemic methicillin resistant Staphylococcus aureus in 1993. , 1994, Communicable disease report. CDR weekly.

[26]  W. Quint,et al.  Concordant clonal delineation of methicillin-resistant Staphylococcus aureus by macrorestriction analysis and polymerase chain reaction genome fingerprinting , 1994, Journal of clinical microbiology.

[27]  R. Gaynes,et al.  An overview of nosocomial infections, including the role of the microbiology laboratory , 1993, Clinical Microbiology Reviews.

[28]  J. Richardson,et al.  Characterization of a strain of methicillin-resistant Staphylococcus aureus (EMRSA-15) by conventional and molecular methods. , 1993, The Journal of hospital infection.

[29]  J. Musser,et al.  Clonal analysis of methicillin-resistant Staphylococcus aureus strains from intercontinental sources: association of the mec gene with divergent phylogenetic lineages implies dissemination by horizontal transfer and recombination , 1992, Journal of clinical microbiology.

[30]  M. Motomiya,et al.  Comparison of a polymerase chain reaction assay and a conventional microbiologic method for detection of methicillin-resistant Staphylococcus aureus , 1992, Antimicrobial Agents and Chemotherapy.

[31]  T. Pennington,et al.  Characterization of Neisseria meningitidis isolates by ribosomal RNA gene restriction patterns and restriction endonuclease digestion of chromosomal DNA , 1991, Epidemiology and Infection.

[32]  S. Sawyer Statistical tests for detecting gene conversion. , 1989, Molecular biology and evolution.

[33]  R. Schleif,et al.  Size fractionation of double-stranded DNA by precipitation with polyethylene glycol. , 1975, Nucleic acids research.