Meticillin-sensitive S. Aureus Research Articles

The distribution of pathogenic and toxigenic genes among MRSA and MSSA clinical isolates

Staphylococcus aureus (S. aureus) is considered as a notorious nosocomial pathogen among hospitalized patients and community-dwelling subjects. Its increasing morbidity and mortality is believed to be due to antibiotic resistance. However, the data concerning molecular properties of infecting strains are few.In this study, a total of 192 S. aureus strains, including 88 (45.8%) meticillin-sensitive S. aureus (MSSA) and 104 (54.2%) meticillin-resistant S. aureus (MRSA) were recovered from clinical samples. The prevalence of subtypes containing staphylococcal cassette chromosome mec (SSCmec), staphylococcal enterotoxins (SEs), toxic shock syndrome toxin (TSST) and exfoliative toxin was assessed by PCR. Antibiotic susceptibility pattern and vancomycin resistance of each isolate were evaluated by disk diffusion method and micro-dilution method, respectively.9 (2.3%) strains required MIC > 2 mg/l of vancomycin, which significantly increased among multi drug resistant (MDR), MRSA and SCCmec type III strains (p < 0.05). 171 (89%), 140 (72.91%), 7 (3.6), 78 (48.6%), 5 (2.6%), 151 (78.64%), 129 (67.18%), 178 (92.7%) and 15 (7.8%) of 192 isolates harbored mecA, entA, entB, entC, entD, entE, eta, etb and tsst-1 genes, respectively. 31 (16.14%), 5 (2.6%), 95 (49.48%) and 7 (3.64%) of 192 isolates carried SCCmec type I, II, III and IV, respectively. We found a significantly higher rate of MRSA and resistance to all tested antibiotics, except to penicillin G, kanamycin and linezolide among the SCCmec type III class (p < 0.05).According to our findings, MSSA isolates should be taken as seriously as MRSA strains due to the potential presence of broad spectrum virulence factor genes.

The dynamic changes of dominant clones of Staphylococcus aureus causing bloodstream infections in the European region: results of a second structured survey.

Staphylococcus aureus is one of the most important human pathogens and meticillin-resistant S. aureus (MRSA) presents a major cause of healthcare- and community-acquired infections. This study investigated the spatial and temporal changes of S. aureus causing bacteraemia in Europe over a five-year interval and explored the possibility of integrating pathogen-based typing data with epidemiological and clinical information at a European level. Between January 2011 and July 2011, 350 laboratories serving 453 hospitals in 25 countries collected 3,753 isolates (meticillin-sensitive S. aureus (MSSA) and MRSA) from patients with S. aureus bloodstream infections. All isolates were sent to the national staphylococcal reference laboratories and characterised by quality-controlled spa typing. Data were uploaded to an interactive web-based mapping tool. A wide geographical distribution of spa types was found, with some prevalent in all European countries. MSSA was more diverse than MRSA. MRSA differed considerably between countries with major international clones expanding or receding when compared to a 2006 survey. We provide evidence that a network approach of decentralised typing and visualisation of aggregated data using an interactive mapping tool can provide important information on the dynamics of S. aureus populations such as early signalling of emerging strains, cross-border spread and importation by travel.

A comparison of long-term outcomes after meticillin-resistant and meticillin-sensitive Staphylococcus aureus bacteraemia: an observational cohort study

Findings from previous studies have suggested that outcomes after meticillin-resistant Staphylococcus aureus (MRSA) bacteraemia are worse than after meticillin-sensitive S. aureus (MSSA) bacteraemia. We assessed whether patients who had MRSA bacteraemia had a higher risk of death and recurrent infections than those who had MSSA bacteraemia. For this observational cohort study, we assessed data from the microbiology laboratory database at the Royal Perth Hospital (WA, Australia). Data were for all patients who had an episode of MRSA bacteraemia between July 1, 1997, and June 30, 2007, and, by use of a computer-generated randomisation sequence, a randomly selected subgroup of patients who had an episode of MSSA bacteraemia (patients with one or more set of blood cultures positive for S. aureus). The primary outcomes were survival time and subsequent infection-related hospital readmissions, analysed by Cox proportional hazards regression with adjustment for important prognostic factors. Of the 583 patients who had an episode of MRSA or MSSA bacteraemia, we used data for the 582 who had complete data linkage: 185 patients who had MRSA bacteraemia and 397 patients who had MSSA bacteraemia. The crude survival time of patients after MRSA bacteraemia was shorter than it was for patients with MSSA bacteraemia (14 months [IQR 1-86] vs 54 months [3-105]; hazard ratio 1·46, 95% CI 1·18-1·79; p=0·01). The adverse association between MRSA and all-cause mortality (0·98, 0·77-1·30; p=0·87) or infection-related mortality (1·22, 0·89-1·69; p=0·22) were not statistically significant after adjustment for important prognostic factors including age, comorbidities, severity of acute illness, metastatic infections, and long-term care facility resident status. After adjustment for these confounding factors, we saw no difference in infection-related hospital readmissions between patients who had MRSA bacteraemia and those who had MSSA bacteraemia (odds ratio 0·95, 95% CI 0·59-1·53; p=0·83). Long-term outcomes after MRSA bacteraemia were worse than those after MSSA bacteraemia through its confounding associations with other prognostic factors. Our findings might have implications for management strategies to control MRSA colonisation. The Medical Research Foundation of Royal Perth Hospital.

Expression of toxic shock syndrome toxin-1 by epidemic meticillin-resistant Staphylococcus aureus 16

BackgroundStaphylococcal toxic shock syndrome (TSS) is a fatal illness attributed to Staphylococcus aureus toxic shock syndrome toxin-1 (TSST-1), encoded by the tst gene. No epidemiological data on TSS exist from the UK. Strains of S aureus may carry the tst gene but not cause TSS, such as the UK strain epidemic meticillin-resistant SA-16 (EMRSA-16). The aim of this study was to determine the epidemiology of TSS in the UK and investigate the expression of TSST-1 by EMRSA-16 and tst+ meticillin-sensitive S aureus (MSSA). MethodsCases of TSS were identified at the UK Health Protection Agency (HPA) and epidemiological features analysed. A selection of clonal complex (CC)-30 EMRSA-16 and TSS-associated MSSA strains was chosen for investigation. The tst gene and promoter region were sequenced in all strains. The staphylococcal pathogenicity island (SaPI) harbouring tst was determined by PCR. Growth of strains in brain heart infusion broth was monitored by optical density (OD600). TSST-1 in supernatants was quantified by western blot using densitometry according to a TSST-1 standard curve. RNA was isolated for tst transcript analysis by quantitative reverse-transcription-PCR (qRT-PCR). Whole genome sequencing was performed on all strains to identify potential mutations in known regulators of toxin synthesis. Findings150 cases of TSS were reported to the HPA over a period of 4 years. Six cases (4%) were attributable to MRSA and the remainder to MSSA. There was no difference in the tst gene or promoter sequence among the strains and tst was carried by SaPI2. TSST-1 production by EMRSA-16 strains commenced earlier in growth than did MSSA strains and was greater in TSS-associated MSSA than in EMRSA-16 strains throughout growth. These results were compared with tst transcription by MSSA and EMRSA16 using qRT-PCR. InterpretationEMRSA-16 produces less TSST-1 than does MSSA of the same lineage; this may be due to differences in global gene regulators, perhaps as a result of changes in PBP2a expression, or the carriage of a large SCCmec element by EMRSA-16. This difference in production may explain the paucity of TSS cases caused by MRSA. FundingUK National Centre for Infection Prevention and Management.

The efficacy of silver dressings and antibiotics on MRSA and MSSA isolated from burn patients

In this study our objectives were (1) to investigate whether meticillin-resistant Staphylococcus aureus (MRSA) showed an increased tolerance to silver wound dressings compared with meticillin-sensitive S. aureus (MSSA); and (2) to evaluate the effects of bacterial phenotypic states of MRSA and MSSA, and pH, on the activity of silver wound dressings and two antibiotics, ampicillin and clindamycin. Twenty MRSA strains and 10 MSSA strains isolated from burns patients in South Africa were evaluated for their susceptibility to a silver alginate and a silver carboxymethyl cellulose wound dressing, employing a corrected zone of inhibition assay, conducted on Mueller Hinton agar and a poloxamer-based biofilm model. When exposed to the two silver dressings, all 30 S. aureus strains showed susceptibility. Possible enhanced antimicrobial efficacy of the silver dressings occurred when pH was lowered to 5.5, compared with a pH of 7.0. When all S. aureus were grown in the biofilm phenotypic state and exposed to both silver dressings and antibiotics, enhanced tolerance was noted. Susceptibility to silver was overall higher for MRSA when compared with MSSA. This study showed that the effect of pH and bacterial phenotypic state must be considered when the antimicrobial activity of silver wound dressings is being investigated. It is evident from the data generated that both pH and the bacterial phenotypic state are factors that induce changes that affect both antimicrobial performance and bacterial susceptibility.

Phagocytosis and intracellular killing of heterogeneous vancomycin-intermediate Staphylococcus aureus strains

Risk factors for invasive infections by heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) may involve resistance to opsonophagocytosis and bacterial killing. hVISA strains typically have a thickened cell wall with altered peptidoglycan cross-linking. To determine whether hVISA may be endowed with an increased resistance to phagocytosis, this study assessed the characteristics of uptake and killing by granulocytes of three hVISA strains. All isolates were analysed by multilocus sequence typing and staphylococcal chromosome cassette mec typing. One of the strains belonged to the Hungarian meticillin-resistant S. aureus (MRSA) clone ST239-MRSA-III and the other two to the New York/Japan MRSA clone ST5-MRSA-II. In the presence of 10 % normal serum, the extent of phagocytosis and killing by blood granulocytes was equivalent for hVISA, MRSA and meticillin-sensitive S. aureus (MSSA) strains. Using granulocytes and serum from one patient who survived hVISA infection, the rate of phagocytosis and killing was also found to be comparable to that by control cells in the presence of 10 % serum. However, phagocytosis and killing of hVISA and MRSA (ATCC 25923) strains by normal granulocytes was markedly decreased in the presence of low concentrations (1 and 2.5 %) of serum from the patient who survived hVISA infection compared with that found with normal human serum. These data suggest that hVISA and MRSA isolates may be more resistant to opsonophagocytosis and bacterial killing than MSSA isolates, at least in some cases.

Increased mortality associated with meticillin-resistant Staphylococcus aureus (MRSA) infection in the Intensive Care Unit: results from the EPIC II study

Controversy continues regarding whether the presence of meticillin resistance increases mortality risk in Staphylococcus aureus infections. In this study, we assessed the role of meticillin resistance in survival of patients with S. aureus infection included in the EPIC II point-prevalence study of infection in critically ill patients performed on 8 May 2007. Demographic, physiological, bacteriological and therapeutic data were collected for 13796 adult patients in 1265 participating Intensive Care Units (ICUs) from 75 countries on the study day. ICU and hospital outcomes were recorded. Characteristics of patients with meticillin-sensitive S. aureus (MSSA) and meticillin-resistant S. aureus (MRSA) infections were compared. Co-morbidities, age, Simplified Acute Physiology Score (SAPS) II, site of infection, geographical region and MRSA/MSSA were entered into a multivariate model, and adjusted odds ratios (ORs) [95% confidence interval (CI)] for ICU and hospital mortality rates were calculated. On the study day, 7087 (51%) of the 13796 patients were classified as infected. There were 494 patients with MRSA infections and 505 patients with MSSA infections. There were no significant differences between the two groups in use of mechanical ventilation or haemofiltration/haemodialysis. Cancer and chronic renal failure were more prevalent in MRSA than in MSSA patients. ICU mortality rates were 29.1% and 20.5%, respectively (P<0.01) and corresponding hospital mortality rates were 36.4% and 27.0% (P<0.01). Multivariate analysis of hospital mortality for MRSA infection showed an adjusted OR of 1.46 (95% CI 1.03–2.06) (P=0.03). In ICU patients, MRSA infection is therefore independently associated with an almost 50% higher likelihood of hospital death compared with MSSA infection.

Simultaneous carriage of multiple genotypes of Staphylococcus aureus in children

The co-existence of multiple genotypes in colonization by Staphylococcus aureus has not been fully investigated. The aim of this study was to evaluate the heterogeneity of S. aureus carriage in children. We evaluated 125 nasal and perianal swab samples that were positive for S. aureus from 76 children scheduled for elective surgery. For each sample, at least four colonies with the same or different morphotypes were selected for analysis. Multiple-locus variable-number tandem-repeat fingerprinting was used to determine the genetic relatedness and to characterize the clonality of the S. aureus strains. Of the 125 swabs, 91 (73 %) contained meticillin-sensitive S. aureus (MSSA), 8 (6 %) contained meticillin-resistant S. aureus (MRSA), and 26 (21 %) contained MSSA and MRSA simultaneously. A total of 738 S. aureus strains were evaluated with a mean of 6 colonies (range 4-15) picked from each culture. Of the 125 swabs, 32 (26 %) samples contained two genetically distinct S. aureus strains and 6 (5 %) contained three different genotypes. Multiple S. aureus strains simultaneously carried by individual children were genetically unrelated to each other. We concluded that the co-existence of multiple genotypes of S. aureus was common. The significance of multiple carriage is yet to be determined, but this intraspecies interplay could be important to pathogenicity and virulence in S. aureus.

Characterisation of methicillin resistant S. aureus strains and risk factors for acquisition in a teaching hospital in northeast India

Purpose: A point prevalence study was carried out in a teaching hospital in Assam to characterise S. aureus strains, establish the rate of colonisation of methicillin resistant S. aureus (MRSA) and associated risk factors for its acquisition. Materials and Methods: Antibiogram-Resistogram profile was done by BSAC standardized disc sensitivity method; Phage and RFLP typing were carried out by the PHLS, London. Results: Single MRSA strain resistant to multiple classes of anti-staphylococcal antibiotics dominated the hospital. The MRSA colonisation rate was found to be 34% (n=29) and 18% (n=80) in orthopaedics and surgery, respectively and only ~1% (n=73) in the medical units. Exposure to ciprofloxacin and surgery were risk factors but duration of hospital stay was not. In contrast, meticillin sensitive S. aureus (MSSA) strains were usually distinct strains and sensitive to most of the anti-staphylococcal antibiotics including 18% to penicillin. Conclusions: The MRSA strain prevalent in the hospital phenotypically resembles the predominant Asian strain viz., Brazilian/Hungarian strains (CC8-MRSA-III). Duration was not a risk factor, which suggests that in absence of exposure to specific antimicrobials, even in a hospital with no or little infection control intervention, a vast majority remain free from MRSA. This underlines the importance of rational prescribing empirical antibiotics.

Biofilm formation by Scottish clinical isolates of Staphylococcus aureus

The biofilm-forming capacity of 972 clinical isolates of Staphylococcus aureus was tested using a high-throughput polystyrene 96-peg plate format. Isolates of S. aureus were collected from patients in hospitals throughout Scotland from 2004 to 2006; 763 of these were meticillin-resistant S. aureus (MRSA) and 209 were meticillin-sensitive S. aureus (MSSA). The biomass of each biofilm was quantified using a crystal violet staining technique. Isolates were divided into those that formed fully established biofilms, moderately attached biofilms and weakly adherent biofilms by comparison with a known biofilm-forming strain. The majority of MRSA (53.8 %) and MSSA (43.5 %) isolates formed moderately attached biofilms. Fully established biofilms were formed by 20.5 % of MRSA isolates and 28.0 % of MSSA isolates, whilst 25.7 % of MRSA isolates and 28.5 % of MSSA isolates formed negligible biofilms. There was no significant correlation between susceptibility to meticillin and biofilm formation (P=0.77). MRSA isolates were divided into clonal types (EMRSA-15, EMRSA-16 and sporadic isolates) based on PFGE genotyping results. EMRSA-15 isolates formed significantly more moderately and fully established biofilms than EMRSA-16 isolates (P<0.001). S. aureus strains isolated from the skin of patients had a significantly greater capacity to form biofilms than isolates from other body sites, including the blood. Microscopic examination of biofilms by scanning electron microscopy (SEM) revealed that poorly adherent biofilm formers failed to colonize the entire surface of the peg, whilst moderately adherent biofilm formers grew in uniform monolayers but failed to develop a mature three-dimensional structure. SEM analysis of an isolate representative of the group that formed fully established biofilms confirmed that this isolate developed a dense biofilm with a textured, multi-layered, three-dimensional structure.

Epidemiology and Economic Impact??of Meticillin-Resistant Staphylococcus aureus

In the past 2 decades, meticillin-resistant Staphylococcus aureus (MRSA) has become an increasingly prevalent problem in healthcare, both in acute care institutions and in the community. MRSA is associated with worse outcomes and higher costs for care than meticillin sensitive S. aureus (MSSA). MRSA is a particular problem in several conditions, including hospital-acquired pneumonia (including ventilator-associated pneumonia), skin and soft tissue infections, and diabetic foot infections. Hospitalisation costs associated with MRSA infection are substantially greater than those associated with MSSA infection, and MRSA has wider economic effects that involve indirect costs to the patient and to society. In several countries, infection control programmes have shown potential economic benefits, as savings accruing from strict and effective control have been shown to outweigh the cost of policy implementation. Standard therapy is based on glycopeptide treatment, usually with vancomycin, although resistance to this agent has emerged. Alternative available treatments for MRSA include teicoplanin, tigecycline, daptomycin, quinupristin-dalfopristin and the oxazolidinone, linezolid, which has a higher acquisition cost than vancomycin but is available as intravenous and oral formulations. Despite some limitations of analyses to date, linezolid has been shown to be cost effective in the treatment of MRSA and appears to be related, in part, to the drug's potential for facilitating earlier discharge from hospital. Current opinion favours rational prescribing to maximise therapeutic benefit and minimise the risk of further antibacterial resistance.

Evaluation of six agglutination tests for Staphylococcus aureus identification depending upon local prevalence of meticillin-resistant S. aureus (MRSA)

Most routine laboratory detection of Staphylococcus aureus isolates is based on rapid agglutination test systems. Failure of agglutination assays to identify meticillin-resistant S. aureus strains (MRSA) has been demonstrated. The aim of this study was to evaluate six commercially available agglutination tests for the detection of meticillin-sensitive S. aureus (MSSA) and mecA-positive MRSA strains. The Dry Spot Staphytect Plus test (Oxoid), the Pastorex Staph Plus test (Bio-Rad), the Slidex Staph-Kit and Slidex Staph Plus test (bioMérieux), the Staphaurex Plus test (Remel) and the Staphylase Test (Oxoid) were used. As determined by pulsed field gel electrophoresis, 52 distinct MRSA strains from five countries, 83 MSSA strains and 150 coagulase-negative staphylococci were included. Species identification and determination of susceptibility patterns were performed using colony morphology, Gram stain, catalase testing, tube coagulase testing, DNase testing, mannitol fermentation, susceptibility testing towards oxacillin by Etest, coagulase gene PCR, fibrinogen receptor gene PCR and PCR of the mecA gene. Sensitivity of the agglutination tests ranged from 82.7 to 100.0 % for MRSA strains and 92.8 to 100.0 % for MSSA strains, respectively. Specificity of the test systems ranged from 91.3 to 99.1 %. None of the six agglutination assays produced correct reactions for all staphylococci tested. Only the Dry Spot Staphytect Plus test correctly identified all 52 MRSA strains. For the other tests kits, sensitivity of MRSA detection was lower than for MSSA isolates. Depending upon the local MRSA prevalence and the parameter of interest (sensitivity or specificity), these test systems may be useful for routine diagnostic purposes.