Molecular epidemiology of methicillin-resistant and methicillin-susceptible Staphylococcus aureus isolates from global clinical trials

Determining the genetic characteristics of Staphylococcus aureus is important for better understanding of the global and dynamic epidemiology of this organism as we witness the emergence and spread of virulent and antibiotic-resistant clones. We genotyped 292 S. aureus isolates (105 methicillin resistant and 187 methicillin susceptible) using a combination of pulsed-field gel electrophoresis, multilocus sequence typing, and SCCmec typing. In addition, S. aureus isolates were tested for the presence of the Panton-Valentine leukocidin (PVL) genes. Isolates were recovered from patients with uncomplicated skin infections in 10 different countries during five phase III global clinical trials of retapamulin, a new topical antibiotic agent. The most common methicillin-resistant clone had multilocus sequence type 8, pulsed-field type USA300, and SCCmec type IV and possessed the PVL genes. This clone was isolated exclusively in the United States. The most common PVL-positive, methicillin-susceptible clone had multilocus sequence type 121 and pulsed-field type USA1200. This clone was found primarily in South Africa and the Russian Federation. Other clones were found at lower frequencies and were limited in their geographic distribution. Overall, considerable genetic diversity was observed within multilocus sequence type clonal complexes and pulsed-field types.     

Changing molecular epidemiology of methicillin-resistant Staphylococcus aureus bloodstream isolates from a teaching hospital in Northern Taiwan

A changing molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) bloodstream isolates from a university-affiliated hospital in Taiwan during a 4-year interval was demonstrated. The changing epidemiology is due to the introduction of a new epidemic clone (sequence type 5) and a local community clone (sequence type 59) of MRSA into the hospital.     

Effect of duplicate isolates of methicillin-susceptible and methicillin-resistant Staphylococcus aureus on antibiogram data

Duplicate Staphylococcus aureus isolates were analyzed to determine the impact of multiple isolates from the same patient on annual antibiogram data. During a 6-year period (1996 to 2001), 3,227 patients with 4,844 S. aureus isolates were evaluated. A total of 39% of patients with methicillin-resistant S. aureus (MRSA) (n = 860) and 23% of patients with methicillin-susceptible S. aureus (MSSA) (n = 2,367) infections had duplicate isolates. Cumulative data show that 91% of the patients during this 6-year period with duplicate isolates (2 to 13 duplicates/year) did not switch between MSSA and MRSA but retained the original S. aureus strain whether it was MSSA or MRSA. Rates of MRSA were calculated for each year by using all isolates and then eliminating duplicates. The impact of duplicate MRSA and MSSA isolates was evaluated by using the ratio of isolates per patient such that ratios of >1.0 indicate >1 isolate per patient. The 6-year ratio for MRSA was 1.90 isolates/patient, and the ratio for MSSA was 1.35. A significant difference (P < 0.05) was noted in the MRSA rates in 4 of 6 years when duplicate isolates were removed. Common phenotypic antibiogram patterns were compared for all MRSA isolates during the 6-year period, and 64% were of a single antibiogram phenotype. Eighty-eight percent of patients with duplicate MRSA isolates had phenotypically identical multiple isolates. The rate of MRSA differs when duplicate isolates are removed from the antibiogram data.     

Genotyping and antimicrobial resistance of Staphylococcus aureus isolates from diseased turkeys

Staphylococcus aureus is a highly versatile pathogen in a large number of domestic animals, including avian species. To gain deeper insight into the epidemiology and diversity of S. aureus associated with articular disease in domestic turkeys, isolates were collected from infected foot joints of turkeys in Brittany (France). A total of 34 isolates were recovered and characterized by means of antimicrobial resistance, staphylococcal protein A typing, macrorestriction pulsed-field gel electrophoresis and micro-array analysis. Thirty isolates were identified as clonal complex (CC) 398 and methicillin-susceptible S. aureus (MSSA), one was identified as a methicillin-resistant S. aureus (MRSA) CC398 isolate, and the remaining were also MSSA and belonged to CC5, CC101, and CC121. Eleven different antimicrobial resistance patterns were detected, with most isolates resistant to penicillin and tetracycline. Based on all typing methods used, the 34 isolates could be divided into 22 different strains. Results on selected isolates, genotyped using microarrays, indicated a high homogeneity among pathogenic MSSA isolates from turkeys. Moreover, all isolates, except the unique MRSA isolate, carried specific φAvβ prophage avian-niche-specific genes, demonstrating the versatility of S. aureus to adapt to the specific ecological poultry niche.     

Genotypic and phenotypic characterization of methicillin-susceptible Staphylococcus aureus isolates misidentified as methicillin-resistant Staphylococcus aureus by the BD GeneOhm MRSA assay

Twenty-three nasal swab samples that tested positive for methicillin-resistant Staphylococcus aureus (MRSA) on initial testing by the BD GeneOhm MRSA assay (BD-MRSA PCR; BD GeneOhm, San Diego, CA) were culture positive only for methicillin-susceptible S. aureus (MSSA) from an enrichment broth. The 23 recovered isolates were confirmed as MSSA by a variety of phenotypic methods, including the BD Phoenix automated microbiology system (BD Diagnostics, Sparks, MD), oxacillin screening agar (BD Diagnostics), BBL CHROMagar MRSA (BD Diagnostics), and a PBP2' assay (Denka Seiken Co., Tokyo, Japan); susceptibilities were determined by using Mueller-Hinton agar with oxacillin. All were positive by nuc PCR, specific for S. aureus, but negative for mecA with one exception. Isolates were characterized by using multiplex PCR methodology to determine structural types and variants (SCCmec typing); additional PCRs were performed for the detection of the ccr and mec complexes, the junkyard regions as well as the Panton-Valentine leukocidin. Pulsed-field gel electrophoresis was used to determine clonality. One phenotypic MSSA isolate contained an intact SCCmec. Twelve MSSA isolates tested positive for MRSA by the BD-MRSA PCR because of amplification of the mec priming site flanking the SCC insertion point, although these isolates lacked mecA. The 10 remaining isolates were not MRSA and tested as MSSA by phenotypic and genotypic assays. In our patient population, diagnostic and surveillance testing and subsequent infection control practices may be impacted by the frequency of these excision events when using the BD-MRSA PCR for MRSA detection.     

Bacteremia due to clonally derived methicillin-resistant, gentamicin-susceptible isolates and methicillin-susceptible, gentamicin-resistant isolates of Staphylococcus aureus

We report recurrent bacteremia due to mixed infection with two clonally derived isolates of Staphylococcus aureus in a patient with Sezary syndrome. The two isolates, one gentamicin resistant and methicillin susceptible and the other gentamicin susceptible and methicillin resistant, developed by the deletion of the mecA, ant(4')Ia, and aacA-aphD genes from a common gentamicin-resistant and methicillin-susceptible ancestor.     

Methicillin resistance in Staphylococcus aureus infections among patients colonized with methicillin-susceptible Staphylococcus aureus

Objectives

We have noticed that patients colonized with methicillin-susceptible Staphylococcus aureus (MSSA) rarely get methicillin-resistant S. aureus (MRSA) infections. The purpose of this study was to compare the odds of a Staphylococcus aureus (SA) infection being an MRSA infection in MSSA carriers, MRSA carriers and non-carriers of SA.

Molecular pattern and antimicrobial susceptibility of methicillin-resistant Staphylococcus aureus isolates at a teaching hospital in Northern Taiwan.

BACKGROUND AND PURPOSE: Methicillin-resistant Staphylococcus aureus (MRSA) infection has progressively increased worldwide. Knowledge of the specific epidemiological pattern of isolates at individual hospitals is important. METHODS: MRSA bacteremia was diagnosed in a total of 68 patients from January 2002 through December 2003, stratified for drug susceptibility and molecular pattern (staphylococcal cassette chromosome mec element [SCCmec] typing and genotypes). RESULTS: SCCmec-A-positive isolates were found on polymerase chain reaction in 58 patients. The most frequent SCCmec types were III (40 cases) of which less than 5% were susceptible to other beta-lactam antibiotics and most were health care-associated, followed by SCCmec type IV (15 cases), that were demonstrated to be community-acquired. SCCmec type IV MRSA isolates were more likely to be susceptible to ciprofloxacin (93.3%), gentamicin (46.7%) and trimethoprim-sulfamethoxazole (93.3%) than type III isolates. All MRSA isolates were susceptible to glycopeptides and vancomycin (minimum inhibitory concentrations <2 microg/mL). Pulsed-field gel electrophoresis with SmaI digestion was used to fingerprint these isolates. A total of 9 genotypes with 26 type-subtypes were identified. Genotype A was the most frequent (9 subtypes) indicating that it is epidemic in this hospital. CONCLUSION: After analysis, SCCmec typing could be used to predict drug susceptibility. Specific clones of S. aureus are circulating in hospital and communities in Taiwan.     

Molecular typing of methicillin-susceptible Staphylococcus aureus isolates collected in the Yogyakarta area in Indonesia, 2006

The characterization of 62 community-associated methicillin-sensitive Staphylococcus aureus (MSSA) isolates from 440 individuals in the Yogyakarta area of Indonesia in 2006 showed that: (i) almost half of the isolates were associated with methicillin-resistant S. aureus lineages [clonal complex (CC)1, CC8 and CC45] and (ii) ten Panton–Valentine leukocidin-positive isolates were associated with CC1 ( n  = 7), CC30 ( n  = 1) and CC51 ( n  = 2). The high Panton–Valentine leukocidin prevalence (16%) among S. aureus is of concern because these strains can cause severe infections and the introduction of staphylococcal cassette chromosome mec into virulent and epidemic MSSA could pose a serious public health threat.     

Resistance to linezolid in a methicillin-susceptible Staphylococcus aureus clinical isolate without previous exposure to oxazolidinones

A linezolid-resistant, methicillin-susceptible isolate of Staphylococcus aureus was obtained from an infected surgical wound in an ambulatory patient. The isolate belonged to ST125 and was susceptible to all the antibiotics tested except linezolid (MIC 8 μg/ml) and levofloxacin. Linezolid resistance could be ascribed to the presence of the mutation G2576T in 2 of the 23S rRNA alleles. The mutant alleles were stably maintained in the absence of antibiotic selection.     

Human lactoferrin binding in clinical isolates of Staphylococcus aureus

Human lactoferrin (HLf) is an iron-binding protein and a host-defence component at the mucosal surface. Recently, a specific receptor for HLf has been identified on a strain of Staphylococcus aureus associated with toxic shock syndrome. We have looked for the occurrence of 125I-HLf binding among 489 strains of S. aureus isolated from various clinical sources. HLf binding was common among S. aureus strains associated with furunculosis (94.3%), toxic shock syndrome (94.3%), endocarditis (83.3%) and septicaemia (82.8%) and other (nasal, vaginal or ocular) infections (96.1%) with a mean binding (in fmol) of 29.1, 21.9, 16.9, 22.2 and 29.2 respectively; the differences between mean HLf binding values of 29.1-29.2, 21.9-22.2 and 16.9 were significant. Furunculosis-associated (low-invasive or localised) isolates were high-to-moderate binders of HLf; 50% gave positive results at a threshold of greater than 31 fmol of 125I-HLf bound. In contrast, endocarditis-associated (high-invasive or systemic) isolates demonstrated low binding and did not bind 125I-HLf at the above threshold level. S. aureus recognised human or bovine Lf. However, bound 125I-HLf was more effectively inhibited in a dose-dependent manner by unlabelled bovine Lf than by homologous HLf. Binding of 125I-HLf to staphylococci was optimal with organisms grown in agar compared with those from broth cultures. The binding capacity of S. aureus was abolished when strains were grown on carbohydrate- and salt-rich agar media. HLf-binding ability of S. aureus did not correlate with fibronectin, fibrinogen, immunoglobulin G or laminin binding.     

Collagen binding in clinical isolates of Staphylococcus aureus.

Collagen binding was examined in 90 strains of Staphylococcus aureus derived from patient samples. Slightly under one-half (39 of 90) of the S. aureus strains bound collagen. Collagen binding in S. aureus did not correlate with either immunoglobulin G or fibronectin binding by these organisms. Chi-square analysis of isolates obtained from patients with complicated bacteremia (bacteremia associated with deep tissue infection) compared with isolates from patients with uncomplicated bacteremia (bacteremia without other infection) showed that the former strains were significantly more likely to have collagen-binding ability. Subcloning of primary isolates from patients with bacteremia showed that all clones from individual patients were either all positive for collagen binding or all negative, suggesting a common clonal origin for this characteristic. The ability to bind collagen could not be induced in strains lacking collagen affinity by repeated subculture in media supplemented with collagen.     

Increased vancomycin MICs for Staphylococcus aureus clinical isolates from a university hospital during a 5-year period

Staphylococcus aureus is one of the most commonly isolated organisms in nosocomial infections. While the prevalence of methicillin-resistant S. aureus (MRSA) continues to increase worldwide, there is concern about an increase in vancomycin MICs among S. aureus strains. The prevalence of MRSA and vancomycin MIC trends in S. aureus from patients in a university hospital were analyzed. Clinical Laboratory Standards Institute (CLSI, formerly NCCLS) reference broth microdilution MIC testing was performed on all clinically relevant S. aureus isolates from January 2000 through December 2004. A total of 6,003 S. aureus isolates were analyzed. No vancomycin-resistant S. aureus isolates were detected. One MRSA isolate had a vancomycin MIC of 8 mug/ml and was confirmed as vancomycin-intermediate S. aureus. Among the 6,002 remaining isolates, a shift in vancomycin MICs from 相似文献   

Acapsular clinical Staphylococcus aureus isolates lack agr function

Staphylococcus aureus is a major human pathogen causing community- and hospital-acquired infections. Capsule production of S. aureus confers protection against host defence. There is a lack of information concerning the association of capsular polysaccharide (CP) expression and activity of the accessory gene regulator (agr) in clinical S. aureus isolates. Production of CP and agr expression were assessed in 195 S. aureus isolates from infected patients at a German University Hospital. Northern blot analysis revealed that S. aureus strains with a non-functional agr locus were more likely to be CP-negative than strains with a functional agr locus.     

Molecular characterization of resistance to mupirocin in methicillin-susceptible and -resistant isolates of Staphylococcus aureus from nasal samples

A total of 15 of 101 (14.8%) nasal methicillin-resistant Staphylococcus aureus (MRSA) isolates exhibited mupirocin resistance (Mup(r)) compared with 1 of 154 (0.6%) methicillin-susceptible Staphylococcus aureus isolates. A total of 14 (93%) isolates exhibiting high-level Mup(r) belonged to a single clone. Horizontal plasmid transfer and transmission of Mup(r) strains contribute to a high incidence of Mup(r) MRSA at our institution.     

Dynamics of methicillin-resistant Staphylococcus aureus and methicillin-susceptible Staphylococcus aureus carriage in pig farmers: a prospective cohort study

Our purpose was to determine the dynamics of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) carriage and its determinants in persons working at pig farms, in order to identify targets for interventions. This prospective cohort study surveyed 49 pig farms in the Netherlands on six sampling dates in 1 year (2010-11). Nasal and oropharyngeal swabs were collected, as well as environmental surface samples from stables and house. Of 110 pig farmers, 38% were persistent MRSA nasal carriers. The average cross-sectional MRSA prevalence was 63%. Methicillin-susceptible S. aureus (MSSA) nasal carriage was associated with fewer MRSA acquisitions (prevalence rate (PR) = 0.47, p 0.02). In multivariate analysis, an age of 40-49 years (PR = 2.13, p 0.01), a working week of ≥40 h (PR=1.89, p 0.01), giving birth assistance to sows (PR=2.26, p 0.03), removing manure of finisher pigs (PR=0.48, p 0.02), and wearing a facemask (PR = 0.13, p 0.02) were significantly related with persistent MRSA nasal carriage. A higher MRSA exposure in stables was associated with MRSA in pig farmers (p <0.0001). This study describes a very high prevalence of LA-MRSA carriage in pig farmers, reflecting extensive exposure during work. We identified the possible protective effects of MSSA carriage and of continuously wearing a facemask during work.     

Prevalence and genetic relatedness of methicillin-susceptible Staphylococcus aureus isolates detected by the Xpert MRSA nasal assay

Methicillin-susceptible Staphylococcus aureus (MSSA) isolates lacking mecA yet testing positive on the Xpert MRSA assay were recovered from culture for 7.7% of 248 Xpert-positive nasal samples. These "false-positive" Xpert results may be attributed to staphylococcal cassette chromosome (SCC) elements without the mecA gene. Pulsed-field gel electrophoresis (PFGE) analysis revealed a diverse population of MSSA strains.     

Molecular genotyping of methicillin-resistant Staphylococcus aureus strains in a teaching hospital in Turkey

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major causes of nosocomial infections in our hospital. Therefore, we aimed to characterize MRSA isolates phenotypically from patients with nosocomial infections at Cumhuriyet University Hospital between December, 1999, and June, 2001, in Sivas by analysis of antibiotic patterns and genotypically using pulsed-field gel electrophoresis (PFGE) and repetitive element sequence-based polymerase chain reaction (rep-PCR). Forty-three nosocomial isolates were collected from various wards. All isolates were resistant to penicillin, tetracycline, oxacillin, and gentamicin. By rep-PCR and by separation of SmaI fragments of genomic DNA using PFGE, one major type (eight subtypes with PFGE) was identified among the strains. This clone was found to be different than some clones such as Iberian, Brazilian, and a major clone that was found in another Turkish University Hospital in Ankara. According to our results, there is a major MRSA clone with a potential to spread in our hospital. Infection control measures should be directed toward restricting the further spread of this clone. Therefore, in accordance with these findings, a surveillance culturing program should be established.