scholarly journals Coagulase gene variants associated with distinct populations of Staphylococcus aureus

2003 ◽  
Vol 130 (2) ◽  
pp. 207-219 ◽  
Author(s):  
P. E. CARTER ◽  
K. BEGBIE ◽  
F. M. THOMSON-CARTER
Keyword(s):  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Multilocus Sequence Typing ◽  
Clonal Complex ◽  
Protein A ◽  
Pulsed Field Gel Electrophoresis ◽  
Gene Variants ◽  
Pfge Analysis ◽  
Pulsed Field ◽  
Clonal Distribution

An identifying characteristic of Staphylococcus aureus is the production of staphylocoagulase (coagulase). The aim of this study was to determine the clonal distribution of coagulase gene (coa) variants within populations of S. aureus defined by multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and protein A variation. The N-terminal region of the coa gene from 43 methicillin-susceptible (MSSA) and 252 methicillin-resistant (MRSA) S. aureus human isolates and 9 animal S. aureus isolates was amplified and digested with HinfI. Twelve types were identified amongst the MSSA isolates and the majority (93%) of MRSA isolates were assigned to 5 of the 12 types. MLST and PFGE analysis identified epidemic populations of MRSA and each epidemic population was characterized by a different coagulase type. Nine of the 12 MLST-defined clonal complex ancestral genotypes recently described each carried a different coagulase type suggesting that coagulase evolution and the evolution of the clonal complexes are intimately related.

scholarly journals Clonal Distribution of Methicillin-Resistant Staphylococcus aureus in Poland

1998 ◽  
Vol 36 (12) ◽  
pp. 3532-3539 ◽  
Author(s):  
T. Leski ◽  
D. Oliveira ◽  
K. Trzcinski ◽  
I. Santos Sanches ◽  
M. Aires de Sousa ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Methicillin Resistance ◽  
Pulsed Field Gel Electrophoresis ◽  
Hospital Environment ◽  
Type I ◽  
Methicillin Resistant ◽  
Pulsed Field ◽  
Clonal Distribution ◽  
Heterogeneous Expression

We report on a study of 158 methicillin-resistantStaphylococcus aureus (MRSA) clinical isolates obtained from 1990 to 1996 in 18 different hospitals in Poland. All isolates were recovered from infection and carriage sites of patients, carriage sites of health care personnel, and hospital environment samples. Fifty-seven MRSA strains described here were studied previously and these were divided into two different clusters according to the degree of heterogeneity of methicillin resistance expression. The aim of this study was to extend the correlation between the two clusters and identify the clonal identities among all isolates by a combination of different methodologies: (i) analysis of mecA polymorphs and Tn554 insertion patterns and (ii) determination of pulsed-field gel electrophoresis patterns of chromosomalSmaI digests. Ninety-seven of 158 strains showed a heterogeneous expression of resistance to methicillin. Among these, 75 (77.3%) were ClaI-mecA type I,ClaI-Tn554 type NH (NH, no homology with transposon Tn554), and pulsed-field gel electrophoresis (PFGE) pattern A (I::NH::A); 10 isolates were III::B::M (10.3%); and the remaining clones included a few or single isolates. The isolates with homogeneous expression of resistance to methicillin (n = 61) were predominantly ClaI-mecA type III (49 of 61 [80.3%]) but had great variability in theirClaI-Tn554 and PFGE patterns. This study confirmed the existence of two main clusters of MRSA in Poland.

Emergence and Characterization of Foodborne Methicillin-Resistant Staphylococcus aureus in Korea

2010 ◽  
Vol 73 (12) ◽  
pp. 2285-2290 ◽  
Author(s):  
CHAE HONG RHEE ◽  
GUN-JO WOO
Keyword(s):  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Multilocus Sequence Typing ◽  
Food Samples ◽  
Pulsed Field Gel Electrophoresis ◽  
Methicillin Resistant ◽  
Pulsed Field ◽  
Fish Samples ◽  
Toxic Shock Syndrome Toxin ◽  
Mrsa Strains

A total of 165 Staphylococcus aureus strains, isolated from different food samples between 2003 and 2006, were tested for antimicrobial susceptibility. The mecA-positive methicillin-resistant S. aureus (MRSA) strains were further characterized by testing for various virulence genes and by molecular typing with multilocus sequence typing and pulsed-field gel electrophoresis. Of the 165 S. aureus isolates, 150 strains (90.9%) were resistant to at least one antibiotic while no strain was resistant to vancomycin. Four strains were resistant to both oxacillin and cefoxitin and were mecA positive. The mecA-positive MRSA strains were isolated from raw meat and fish samples (two beef samples and two fish samples) and were resistant to β-lactam antibiotics. Based on multilocus sequence typing analysis, the isolates were assigned to sequence type 1 (ST1), ST72, and an undetermined ST (ST72 slv). All four MRSA isolates were shown to be enterotoxigenic. The ST1 MRSA isolate harbored the sea-seh gene combination and the ST72 and ST72 slv MRSA strains harbored the seg-sei and the sea-seg-sei gene combinations, respectively. However, none of the MRSA isolates had the genes for Panton-Valentine leukocidin, toxic shock syndrome toxin 1, and exfoliative toxins. The pulsed-field gel electrophoresis patterns of the ST72 isolates in our study were highly similar, even though they were isolated from food samples in different years and from different regions of Korea.

scholarly journals Comparison of Protein A Gene Sequencing with Pulsed-Field Gel Electrophoresis and Epidemiologic Data for Molecular Typing of Methicillin-Resistant Staphylococcus aureus

2000 ◽  
Vol 38 (4) ◽  
pp. 1347-1351 ◽  
Author(s):  
Yi-Wei Tang ◽  
Michael G. Waddington ◽  
Douglas H. Smith ◽  
Janice M. Manahan ◽  
Peggy C. Kohner ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Sequence Analysis ◽  
Molecular Typing ◽  
Gel Electrophoresis ◽  
Protein A ◽  
Pulsed Field Gel Electrophoresis ◽  
Sequencing Analysis ◽  
Methicillin Resistant ◽  
Pulsed Field ◽  
Epidemic Period

The epidemiologic relatedness of methicillin-resistantStaphylococcus aureus (MRSA) isolates is currently determined by analysis of chromosomal DNA restriction patterns by pulsed-field gel electrophoresis (PFGE). We have evaluated an alternative typing system (MicroSeq StaphTrack Kit; Perkin-Elmer Biosystems) based on the sequence analysis of the chromosomally encoded polymorphic repeat X region of the S. aureus protein A (spa) gene. A total of 69 clinical MRSA isolates were divided into 18 groups according to the number and nucleotide sequences of the spa repeats. Molecular typing results obtained both by spa sequencing and from the PFGE patterns were concordant except for one group, which contained 20 isolates recovered over a 2-year period from hospitalized patients at the Mayo Clinic. Although the spa typing patterns were indistinguishable for those isolates, PFGE analysis yielded seven related but distinguishable patterns. Further coagulase gene sequence analysis subtyped those 20 strains into four groups which followed distinct temporal and geographic distributions. During a 2-year epidemic period there were up to 7 fragment changes in PFGE patterns among epidemiologically related isolates, suggesting that PFGE may be unsuitable for long-term typing of strains involved in epidemics. Although more limited than PFGE in discriminatory power, spa sequencing analysis could be used as a screening method for typing of MRSA strains because of the shorter turnaround time, ease of use, and the inherent advantages of sequence analysis, storage, and sharing of information.

scholarly journals High Heterogeneity within Methicillin-Resistant Staphylococcus aureus ST398 Isolates, Defined by Cfr9I Macrorestriction-Pulsed-Field Gel Electrophoresis Profiles and spa and SCCmec Types

2009 ◽  
Vol 76 (3) ◽  
pp. 652-658 ◽  
Author(s):  
M. A. Argudín ◽  
A. Fetsch ◽  
B.-A. Tenhagen ◽  
J. A. Hammerl ◽  
S. Hertwig ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Protein A ◽  
Pulsed Field Gel Electrophoresis ◽  
Clinical Samples ◽  
Methicillin Resistant ◽  
Pulsed Field ◽  
Outbreak Investigations ◽  
Type V ◽  
Sccmec Types

ABSTRACT During recent years, the animal-associated methicillin-resistant Staphylococcus aureus clone ST398 has extensively been studied. The DNA of these isolates turned out to be refractory to SmaI restriction, and consequently, SmaI is unsuitable for subtyping this clone by standard pulsed-field gel electrophoresis (PFGE). Very recently, ST398 DNA was shown to be digested by Cfr9I, a neoschizomer of SmaI. In the present study, we employed Cfr9I PFGE on 100 German and 5 Dutch ST398 isolates and compared their PFGE profiles, protein A gene variable repeat regions (spa types), and types of the staphylococcal cassette chromosome mec (SCCmec). The isolates (from healthy carrier pigs, clinical samples from pigs, dust from farms, milk, and meat) were assigned to 35 profiles, which were correlated to the SCCmec type. A dendrogram with the Cfr9I patterns assigned all profiles to two clusters. Cluster A grouped nearly all isolates with SCCmec type V, and cluster B comprised all SCCmec type IVa and V* (a type V variant first identified as III) carriers plus one isolate with SCCmec type V. Both clusters also grouped methicillin-susceptible S. aureus isolates. The association of the majority of isolates with SCCmec type V in one large cluster indicated the presence of a successful subclone within the clonal complex CC398 from pigs, which has diversified. In general, the combination of Cfr9I PFGE with spa and SCCmec typing demonstrated the heterogeneity of the series analyzed and can be further used for outbreak investigations and traceability studies of the MRSA ST398 emerging clone.

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