Diversity among community isolates of methicillin-resistant Staphylococcus aureus in Australia

Community methicillin-resistant Staphylococcus aureus (CMRSA) strains are being isolated with increasing frequency around the world. In Western Australia CMRSA are endemic in geographically remote communities and have been found to belong to five different contour-clamped homogeneous electric field (CHEF) electrophoretic patterns. Representatives of each of these CHEF patterns have been compared to CMRSA representative of CHEF patterns from other Australian states and New Zealand. With one exception, all of the isolates were nonmultiresistant and were not resistant to many antimicrobial agents other than the beta-lactams. With one exception, which is not believed to be a CMRSA, all of the isolates harbored a beta-lactamase plasmid. Erythromycin resistance was associated with a 2-kb plasmid. One of the beta-lactamase plasmids was found to be able to acquire additional resistance determinants to become a multiple resistance plasmid. There were 10 multilocus sequence types belonging to eight distantly related clonal complexes of S. aureus. One new sequence type was found. Although most of the CMRSA harbored the type IVa SCCmec, a type IV structural variant was found and two new SCCmec types were identified. Protein A gene (spa) typing revealed two new spa types and, with two exceptions, corresponded to multilocus sequence typing. In contrast to other reports on CMRSA, most of the CMRSA strains studied here did not contain the Panton-Valentine leukocidin genes. The results also demonstrate that nonmultiresistant hospital strains such as UK EMRSA-15 may be able to circulate in the community and could be mistaken for CMRSA based on their resistance profiles.     

Management of methicillin-resistant Staphylococcus aureus infections

This review addresses selected aspects of the management of severe healthcare-associated infections due to methicillin-resistant Staphylococcus aureus (MRSA), including the limitations of current therapy, potential alternative agents, new therapeutic options, clinical approaches to MRSA bacteraemia/endocarditis and ventilator-associated pneumonia, and strategies to improve outcomes in patients with severe MRSA infections.     

Genetic diversity among methicillin-resistant Staphylococcus epidermidis (MRSE)

We selected 106 methicillin-resistant Staphylococcus epidermidis (MRSE) and 22 methicillin-susceptible S. epidermidis (MSSE) hospital isolates--each with a different PFGE pattern--for more detailed documentation of genetic diversity. The 106 MRSE isolates showed extensive variation in the SmaI DNA fragments hybridizing with the DNA probe for mecA, the molecular size of which varied from as low as 20 kb up to over 500 kb. Parallel variation was also observed in the size of DNA fragments hybridizing with the chromosomal genes orfX and gyrA, and this was also observed in MSSE isolates. In contrast, SmaI fragments associated with the housekeeping genes murE and aroE, both located distantly from orfX, showed little size variation. Typing for the mec complex and ccr identified 10 different SCCmec structures and a large number of strains (21 isolates) that were non-typeable. The majority of strains studied (36%) carried a SCCmec type IV-like structure, including strains with non-related PFGE profiles. On the other hand, closely related strains often carried different types of SCCmec. The findings indicate that the acquisition and/or loss of mobile genetic elements, including various structural types of SCCmec, may occur frequently in the vicinity of the orfX gene on the S. epidermidis chromosome.     

Genetic differentiation of methicillin-resistant Staphylococcus aureus strains from Korea and Japan

In this study, we evaluated genetic differentiation between methicillin-resistant Staphylococcus aureus (MRSA) strains from Korea and Japan. Seventy-five MRSA strains, including 25 h VISA strains, were analyzed by molecular typing methods, including multilocus sequence typing (MLST), SCC mec typing, and spa typing. The most prevalent genotype of MRSA strains, in both Korea and Japan, was ST 5-MRSA-II with the DMGMK spa motif, characteristic of the New York/Japan MRSA clone. In spite of these common features in MRSA strains from Korea and Japan, we also observed some genotypic divergence in MRSA from the two countries. Several spa types might be differentiated from a prevalent prototype (TJMBMDMGMK) that is shared by the two countries, revealing a unique geographic distribution. SCC mec type II lacking pUB110, designated type IIA, was found more frequently in Korea than in Japan. The rate of gentamicin resistance was also dramatically different between the two countries: 87.2% (Korea) vs. 28.6% (Japan). These preliminary findings suggested that MRSA strains from Korea and Japan might have originated from a common ancestor, but then clearly differentiated according to locality. A further comprehensive study should be performed to document the hypotheses from this study.     

Life-threatening community-acquired methicillin-resistant Staphylococcus aureus infection in Australia

Eight patients with invasive bacteremic community-acquired methicillin-resistant Staphylococcus aureus infection in southeast Queensland, Australia, are reported. One patient died of septic shock. Haematogenous seeding to lungs, bone, and other sites was common. All isolates carried the virulence factor Panton-Valentine leukocidin and were either the southwest Pacific clone or the newly described Queensland clone. Clinicians should consider community-acquired methicillin-resistant Staphylococcus aureus infection in any patient presenting to hospital with severe staphylococcal sepsis or pneumonia.     

Lysogenicity of methicillin-resistant strains of Staphylococcus aureus

The lysogenic status of 23 strains of methicillin-resistant Staphylococcus aureus, isolated at the Royal Prince Alfred Hospital, Sydney, since 1980, was studied. Twenty strains, belonging to the four predominant phage types isolated in this hospital, carried the same lysogenic phage which we have designated C. Three other phages were isolated from five strains belonging to phage type 84/85/90. The presence of phage C had little effect on the phage-typing pattern of the strains. Similarly, lysogenization with the other three phages did not result in a significant change in phage-typing patterns. However, when strain 1489, isolated in 1969, was lysogenized with these three phages, there was a change in phage-typing pattern. Lysogenization of this strain with phage 47T resulted in a marked loss of sensitivity to both group-I and group-III phages. The lysogenic status of these methicillin-resistant strains of S. aureus was compared with that of strains isolated between 1967 and 1970. There was no evidence that the strains isolated recently were either related to, or derived from, the earlier ones.     

Genetic diversity in CC398 methicillin-resistant Staphylococcus aureus isolates of different geographical origin

Staphylococcus aureus of sequence type 398 has emerged in Europe, North America and Asia, and has typically been associated with livestock and their human contacts. We analysed two Panton–Valentine leukocidin (PVL)-negative t034-ST398 isolates from humans in contact with pigs and two t034-ST398 PVL-positive isolates from two unrelated, adopted Chinese children, using multistrain microarrays to determine genomic variability between the two sets of isolates. The ST398 isolates clearly belong to the same lineage when compared to other clonal lineages. However, the four isolates cluster into two distinct groups corresponding to differences in epidemiology based on mobile genetic elements and resistance patterns, suggesting that the two groups are epidemiologically distinct.     

Genetic diversity among type emm28 group A Streptococcus strains causing invasive infections and pharyngitis

Genome sequencing of group A Streptococcus (GAS) has revealed that prophages account for the vast majority of gene content differences between strains. Serotype M28 strains are a leading cause of pharyngitis and invasive infections, but little is known about genetic diversity present in natural populations of these organisms. To study this issue, population-based samples of 568 strains from Ontario, Canada; Finland; and Houston, Texas, were analyzed. Special attention was given to analysis of variation in prophage-encoded virulence gene content by a PCR-based method. Thirty and 29 distinct prophage-encoded virulence gene profiles were identified among pharyngitis and invasive infection isolates. Thirteen profiles, representing the majority of the strains, were shared between these two classes of isolates. Significant differences were observed in the frequency of occurrence of certain prophage toxin gene profiles and infection type. M28 strains are highly diverse in prophage-encoded virulence gene content and integration site, supporting the key concept that prophages are critical contributors to GAS genetic diversity and population biology. Nucleotide sequence variation in the emm gene (encodes M protein) was also examined. Only three allelic variants were identified in the hypervariable portion of the emm28 gene. All but one strain had the same inferred amino acid sequence in the first 100 amino acids of the mature M28 protein. In contrast, size differences in the emm28 gene and inferred protein due to variable numbers of C-terminal repeats were common. The presence of macrolide resistance genes (mefA, ermB, and ermTR) was analyzed by PCR, and less than 2% of the strains were positive.     

Genetic variation among methicillin-resistant Staphylococcus aureus isolates from cancer patients in Saudi Arabia

One hundred and twenty methicillin-resistant Staphylococcus aureus (MRSA) isolated from cancer and non-cancer patients in Saudi Arabia were investigated for antibiotic resistance, virulence determinants and genotypes. The majority of MRSA isolates from cancer (n?=?44, 73.3 %) and non-cancer patients (n?=?34, 56.7 %) were multi-resistant to more than four classes of antibiotics. Virulence gene profiling showed that all strains were commonly positive for adhesin genes, except ebps and bbp genes, which were not detected in any isolate. Although the presence of adhesin genes varied slightly among MRSA isolates from cancer and non-cancer patients, these variations were not found to be statistically significant. In contrast, the presence of the toxin genes seb, sec, seg and sei was significantly elevated in MRSA strains isolated from cancer patients. Multilocus sequence typing (MLST) detected six and nine sequence types (STs) among isolates from cancer and non-cancer patients, respectively. Using spa typing, 12 and 25 types were detected, including four new types. The ability of different MRSA clones to become multi-resistant and their ability to acquire different virulence factors may contribute to their success as pathogens in individual groups of patients.     

Undecatungstophosphate sensitized strains of methicillin-resistant Staphylococcus aureus to beta-lactams

Previously, a factor (Factor T) was found in aged mixtures of tungstate and phosphate, which greatly sensitizes strains of methicillin-resistant Staphylococcus aureus to beta-lactams. Factor T was purified and identified as undecatungstophosphate([PW11O39]7-). Undecatungstosilicate([SiW11O39]8-), a compound closely related to undecatungstophosphate, showed a similar enhancing effect. Chemically, these compounds are classified as "polyoxotungstates", and it is expected that these tungsten compounds will be useful as a "tool" in laboratory tests: i.e., in screening media for highly resistant MRSA strains. They may be also useful to investigate the resistant mechanism of the bacterial cells.     

Genetic diversity of community-associated methicillin-resistant Staphylococcus aureus in southern Stockholm, 2000–2005

This study investigated the molecular epidemiology of 104 community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) isolates from southern Stockholm during the period 2000–2005. The isolates were analysed by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing, staphylococcal chromosomal cassette (SCC) mec typing and detection of genes encoding Panton–Valentine leukocidin (PVL). Overall, 28 distinct PFGE patterns and 13 sequence types (STs) were identified. ST80, ST8, ST88 and ST150 were the major CA-MRSA clones in the area, and these accounted for 75% (78/104) of all CA-MRSA isolates. ST150 isolates, which have, to date, been found only in Sweden, were isolated exclusively from a group of homeless individuals. Eighty-six (83%) of the 104 isolates in the study possessed SCC mec IV, found in ten different STs, while 16 isolates possessed SCC mec V. The PVL genes were detected in 56% (58/104) of the isolates. Strain ST80-MRSA-IV carrying PVL genes predominated over the 6-year period and accounted for 38% of all isolates. However, a polyclonal tendency was observed among the CA-MRSA isolates recovered in recent years.     

Countrywide molecular survey of methicillin-resistant Staphylococcus aureus strains in Poland

The present investigation was undertaken to assess the proportion of methicillin-resistant Staphylococcus aureus (MRSA) strains among hospital-acquired isolates and to determine the clones of MRSA currently circulating in Poland by using a number of molecular techniques. Between January and May 2005, methicillin resistance was investigated among a total of 915 S. aureus isolates collected from 39 hospitals. A total of 208 (22.7%) isolates were positive for the mecA gene by PCR. The molecular characterization of MRSA isolates was carried out by the multiple-locus variable-number tandem repeat fingerprinting, pulsed-field gel electrophoresis, multilocus sequence typing, and staphylococcal chromosomal cassette mec (SCCmec) typing methods. The Hungarian (PFGE B; ST239, SCCmec type III [ST239-III]), Iberian (ST247-I), and Berlin (ST45-IV) clones were predominant, representing approximately 52.9, 11.5, and 10.0% of the MRSA isolates, respectively. A decline in the proportion of earlier MRSA clones, such as ST5-IV (a Pediatric clone), ST80-IV) (a Mediterranean clone), ST239-III (a Polish and Brazilian clone), and ST30-IV (a southwest Pacific clone) was observed. Additionally, the emergence of an MRSA clone with SCCmec type V, possibly representing a community-acquired strain, was observed in two hospitals during this study.     

Extreme genetic diversity of methicillin-resistant Staphylococcus epidermidis strains disseminated among healthy Japanese children

For the past few years, we have been observing the dissemination of methicillin-resistant staphylococci in the community. From 2001 to 2003, an evaluation of nasal samples from 1,285 children in five day-care centers and two kindergartens in three districts in Japan revealed that methicillin-resistant coagulase-negative staphylococci (MRC-NS) have been widely disseminated in the Japanese community. Their prevalence is much greater than community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA). Forty-nine children (3.81%) were colonized with MRSA, whereas 390 children (30.35%) were colonized with MRC-NS. These MRC-NS strains predominantly harbored a pair of cassette chromosome recombinase types A2 and B2 (ccrAB2). Of these, 40.8% harbored type IVa staphylococcal cassette chromosome mec (SCCmec) elements, a distinct/characteristic type of SCCmec in pandemic clones of CA-MRSA. Interestingly, there was also a high frequency of nontypeable strains which possessed atypical structures compared to previous SCCmec types. Among the MRC-NS, the majority of strains (63.59%) were methicillin-resistant Staphylococcus epidermidis (MRSE). Their genotypes, as judged from pulsed-field gel electrophoresis (PFGE), were highly diverse. They were so diverse that there was no sign of an immediate transmission of any MRSE clone among children in the same institutions. In a previous report, we expounded that a few CA-MRSA clones with distinct SCCmec types were disseminated among children in the same institutions. Au contraire, with the case of CA-MRSE, there was no single genotype of CA-MRSE disseminated among children even in the same institution or class.     

Nasal carriage of Staphylococcus aureus and its influence on hospital infections caused by methicillin-resistant strains

A short review of the literature on the actual epidemiological situation related to nosocomial infections caused by methicillin-resistant Staphylococcus aureus and the importance of the nasal carriage of this pathogen is given together with selected data derived from our own studies on these problems.     

Recurrent community-acquired methicillin-resistant Staphylococcus aureus infections in an HIV-infected person

HIV-infected persons are at heightened risk for recurrent community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infections, but there are limited data regarding the molecular characterization of these events. We describe an HIV-infected patient with 24 soft tissue infections and multiple colonization events. Molecular genotyping from 33 nonduplicate isolates showed all strains were USA300, Panton-Valentine leukocidin (PVL) and arginine catabolic mobile element (ACME) positive, and genetically related.