Sporadic Methicillin-resistant Staphylococcus Aureus Research Articles

Successful control of a methicillin-resistant Staphylococcus aureus outbreak in a burn intensive care unit by addition of universal decolonization with intranasal mupirocin to basic infection prevention measures

Methicillin-resistant Staphylococcus aureus (MRSA) is frequently implicated in health care-associated outbreaks in burn intensive care units, incurring substantial morbidity and mortality to these high-risk patients and excess costs to health care systems. MRSA health care-associated infections (HAIs) were noted before and after the implementation of basic infection prevention measures and the subsequent implementation of universal decolonization with intranasal mupirocin. Pulsed-field gel electrophoresis was used to determine the relatedness of clinical isolates. A case-control study was conducted to characterize the risk factors for MRSA HAIs. Basic interventions failed to decrease the rate of MRSA HAIs, although compliance with these interventions was high throughout the study. MRSA HAIs decreased from 8.53 HAIs per 1,000 patient days before the implementation of intranasal mupirocin to 3.61 HAIs per 1,000 patient days after the implementation of intranasal mupirocin (P = .033). Pulsed-field gel electrophoresis disclosed 10 unique clones with no large clusters. The case-control study revealed a significant association between MRSA HAIs and lengths of stay, body surface area burned, intubation, pressor requirement, leukocytosis, lactic acidosis, development of pneumonia, MRSA colonization, and death. Basic environmental and behavioral interventions fell short of controlling a low-count, sporadic, and multiclonal MRSA outbreak in the burn intensive care unit of a tertiary medical center. However, the added implementation of universal decolonization with intranasal mupirocin was effective. Burn victims with greater disease severity were at higher risk for MRSA HAIs.

Antimicrobial resistance and molecular analysis of methicillin-resistant Staphylococcus aureus collected in a Spanish hospital.

Clonal distribution of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals may differ according to the geographic location and time period. Knowledge of MRSA clonal epidemiology in hospital settings involves much more than the study of healthcare-associated MRSA (HA-MRSA) clones. In recent years, investigators have documented the introduction of both community-associated MRSA (CA-MRSA) and livestock-associated MRSA (LA-MRSA) clones, the emergence of clones carrying Staphylococcal cassette chromosome mec (SCCmec) XI, and the genetic diversity among sporadic MRSA isolates. The allocation of certain antibiotypes to dominant MRSA clones in an institution allows their use as phenotypic markers for a preliminary search for new clones, early detection of clonal shift, and as a guide for better empirical therapy, infection control, and treatment within a particular institution. For these reasons, we identified 938 strains detected in a System of Universal Active Surveillance of MRSA in clinical samples during the period 2009-2010, obtaining the clonal distribution of MRSA at the Hospital Universitario de Canarias (Tenerife, Spain) and the relationship between antimicrobial susceptibility and three major clones present. The antibiotypes that best defined the ST5-MRSA-IV (Pediatric) clone showed resistance to tobramycin and susceptibility to clindamycin, erythromycin, gentamicin, rifampin, trimethoprim-sulfamethoxazole, vancomycin, quinupristin/dalfopristin, and linezolid, whereas the ST22-MRSA-IV clone (EMRSA-15) showed susceptibility to these antibiotics, and finally, the ST36-MRSA-II clone (EMRSA-16) was resistant to clindamycin, erythromycin, and tobramycin and susceptible to the remaining antimicrobials. Similar observations would allow the early detection of changes in clonal epidemiology by analysis of antimicrobial susceptibility of the isolates within a single institution.

Extensive Genetic Diversity Identified among Sporadic Methicillin-Resistant Staphylococcus aureus Isolates Recovered in Irish Hospitals between 2000 and 2012

Clonal replacement of predominant nosocomial methicillin-resistant Staphylococcus aureus (MRSA) strains has occurred several times in Ireland during the last 4 decades. However, little is known about sporadically occurring MRSA in Irish hospitals or in other countries. Eighty-eight representative pvl-negative sporadic MRSA isolates recovered in Irish hospitals between 2000 and 2012 were investigated. These yielded unusual pulsed-field gel electrophoresis and antibiogram-resistogram typing patterns distinct from those of the predominant nosocomial MRSA clone, ST22-MRSA-IV, during the study period. Isolates were characterized by spa typing and DNA microarray profiling for multilocus sequence type (MLST) clonal complex (CC) and/or sequence type (ST) and SCCmec type assignment, as well as for detection of virulence and antimicrobial resistance genes. Conventional PCR-based SCCmec subtyping was undertaken when necessary. Extensive diversity was detected, including 38 spa types, 13 MLST-CCs (including 18 STs among 62 isolates assigned to STs), and 25 SCCmec types (including 2 possible novel SCCmec elements and 7 possible novel SCCmec subtypes). Fifty-four MLST-spa-SCCmec type combinations were identified. Overall, 68.5% of isolates were assigned to nosocomial lineages, with ST8-t190-MRSA-IID/IIE±SCCM1 predominating (17.4%), followed by CC779/ST779-t878-MRSA-ψSCCmec-SCC-SCCCRISPR (7.6%) and CC22/ST22-t032-MRSA-IVh (5.4%). Community-associated clones, including CC1-t127/t386/t2279-MRSA-IV, CC59-t216-MRSA-V, CC8-t008-MRSA-IVa, and CC5-t002/t242-MRSA-IV/V, and putative animal-associated clones, including CC130-t12399-MRSA-XI, ST8-t064-MRSA-IVa, ST398-t011-MRSA-IVa, and CC6-t701-MRSA-V, were also identified. In total, 53.3% and 47.8% of isolates harbored genes for resistance to two or more classes of antimicrobial agents and two or more mobile genetic element-encoded virulence-associated factors, respectively. Effective ongoing surveillance of sporadic nosocomial MRSA is warranted for early detection of emerging clones and reservoirs of virulence, resistance, and SCCmec genes.

Antidiabetic and antimicrobial activities of fractions and compounds isolated from <i>Berberis brevissima</i> Jafri and <i>Berberis parkeriana</i> Schneid

Methanol extracts of the roots of B. brevissima and B. parkeriana were examined for antidiabetic and antimicrobial activities. Six compounds i.e berberine, 8-oxo-berberine, dehydrocheilanthifoline, columbamine, jatrorrhizine and gluta-mic acid were isolated from the active fractions of these species for the first time. The antidiabetic activity was determined against Protein Tyrosine Phosphatase 1B (PTP 1B), a negative insulin regulator while the antimicrobial activity was performed against four bacterial strains: Mycobacterium marinum ATCC BAA535, sporadic Methicillin Resistant Staphylococcus aureus (SMRSA), endemic Methicillin Resistant Staphylococcus aureus (EMRSA) and Escherichia coli ATCC.8739 at different concentrations. Amongst the isolated compounds 8-oxo-berberine was found to be the most active antidiabetic (29 % of the positive control) while jatrorrhizine was the most active and selective antimicrobial against SMRSA and EMRSA (-1.2% of the DMSO (D) control).

Evidence that contaminated surfaces contribute to the transmission of hospital pathogens and an overview of strategies to address contaminated surfaces in hospital settings

Evidence that contaminated surfaces contribute to the transmission of hospital pathogens comes from studies modeling transmission routes, microbiologic studies, observational epidemiologic studies, intervention studies, and outbreak reports. This review presents evidence that contaminated surfaces contribute to transmission and discusses the various strategies currently available to address environmental contamination in hospitals.

Evaluation of the TPX MRSA assay for the detection of methicillin-resistant Staphylococcus aureus

The aim of this study was to evaluate a new type of assay for the phenotypic detection of methicillin-resistant Staphylococcus aureus (MRSA). The assay is based on a point-of-care compatible two-photon excitation fluorescence detection technology (TPX). A collection of 243 epidemic MRSA isolates was tested in addition to 138 sporadic MRSA and 101 negative control strains. The assay proved to be both sensitive (97.9%) and specific (94.1%) in the identification of MRSA, with adequate positive (98.4%) and negative (92.2%) predictive values. The time required for obtaining a positive test result was less than 14 h for 99.0% of the MRSA true-positive samples. After a test run, the selectively enriched reaction mixtures may be recovered and further studied by molecular or standard phenotypic methods. The main benefits of the TPX methodology include a simple assay procedure, low reagent consumption, and a high-throughput capacity.

Eradication of methicillin-resistant Staphylococcus aureus in a neonatal intensive care unit: Which measures for which success?

Various strategies for controlling methicillin-resistant Staphylococcus aureus (MRSA) outbreaks in neonatal intensive care units (NICUs) have been tried, with varying levels of success. We report a MRSA outbreak occurring between April 2004 and August 2007 in a 24-bed NICU in a large university hospital. We describe the difficulties involved in implementing measures to control the MRSA outbreak and the possible contribution of each measure. Cases were defined as neonates with MRSA obtained from either clinical cultures or surveillance cultures (from the anterior nares). Systematic screening of neonates for colonization was performed only between February and December 2005. Successive control strategies included barrier precaution and isolation in individual rooms, mupirocine ointment for neonates and health care workers, cohort isolation, hand hygiene observation, and staff training. During the routine surveillance culture period (February to December 2005; 48 weeks), 46 neonates were found to be positive for MRSA and were treated with mupirocin. After December 2005, the outbreak was controlled, but the ongoing spread was not eradicated; 9 sporadic MRSA cases were detected by clinical culture up to August 2007. The widespread use of mupirocine in staff and patients did not control the outbreak and is not recommended. The later control appeared to coincide with increased hand hygiene audits and training for staff, along with appropriate cohort isolation of neonates and cohort nursing.

Emergence of SCCmec Type IV as the Most Common Type of Methicillin-Resistant Staphylococcus aureus in a University Hospital

The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) has dramatically changed over the last decade by the emergence of community-associated MRSA (CA-MRSA). Recent studies indicate that these strains have already spread to hospitals. To evaluate if SCCmec type IV and Panton-Valentine leukocidin (PVL) are unambiguous markers of CA-MRSA, we analyzed 77 sporadic MRSA strains isolated, in our low MRSA incidence university hospital, from inpatients between 2000 and 2004. MRSA strains were analyzed by staphylococcal cassette chromosome mmecec (SCCmec) typing, PCR for PVL genes and pulsed-field gel electrophoresis (PFGE). MRSA was classified in HA-MRSA or CA-MRSA according to Centers for Disease Control and Prevention (CDC) criteria. Antimicrobial susceptibility testing was performed using microbroth dilution method following CLSI recommendations. Among 77 sporadic single-patient strains, SCCmec types I-IV and four subtypes were identified. Type IV/IVA was most common (42.9%).The distribution of SCCmec types changed over the years. Type IV/IVA strains increased from 33.3% in 2000 to 57.9% in 2004. Type IV strains were resistant to ciprofloxacin in 81.8%, and in 9.1% to tobramycin while type IVA strains were 100% resistant to both antimicrobials. In contrast, non-type IV/IVA strains were resistant to ciprofloxacin in 86.4%, and in 75.0% to tobramycin. Only one strain was PVL positive and harbored SCCmec type III variant. By PFGE analysis, the 33 SCCmec type IV/IVA strains comprised 12 distinct genotypes. 36.4% of 11 CA-MRSA and 43.9% of 66 HA-MRSA harbored SCCmec type IV/IVA. Type IV/IVA has become the most common SCCmec type in inpatients of our university hospital. The SCCmec type IV/IVA is present in both CA-MRSA and HA-MRSA limiting its use as a marker for CA-MRSA.

부산지역 설사환자에서 분리한 MRSA 균주의 다양성 분석

부산 지역 5개 2차 종합병원 설사환자 변에서 유래한 메치실린 내성 황색포도상구균(MRSA)에 대한 연구 결과, 분리된 황색포도상구균 142주 중 총 49주(34.5%)가 MRSA인 것으로 확인되었으며, 황색포도상구균 중 MRSA 분리율이 '04, '05, '06년에 걸쳐 각각 30.0%, 32.7%, 46.7%인 것으로 나타나, 지속적으로 증가하고 있음을 알 수 있었다. MRSA 균주의 항균제 내성 양상은 75.5%가 10종 이상의 항균제에 내성을 나타내었으며, 특히 11종 항균제에 대한 다제 내성균이 가장 많은 것으로 확인되었다. MRSA주의 장독소 유전자 시험을 실시한 결과 51%가 장독소 유전자를 가지고 있지 않았고, 30.6%가 독소유전자 A를 가지고 있었으며 그 밖에 C (8.3%), B (4.1%) 순이었다. MRSA 총 49주에 대해 PFGE를 수행하여 dendrogram을 작성한 결과, 크게 7개의 그룹으로 구분되었으며, 부산지역 전체에 뚜렷하게 동일한 균주의 유행은 확인 할 수 없었으나, 종합병원 내에 소규모의 MRSA 유행은 있을 것으로 사료되었다. We investigated the molecular epidemiological characteristic of methicillin resistant Staphylococcus aureus (MRSA) strains isolated from stool samples in Busan from 2004 to 2006. Among 142 isolates of S. aureus, 49 isolates (34.5%) were confirmed as MRSA. With the antimicrobial susceptibility tests, 37 isolates (75.5%) showed multiple resistance to more than 10 antibiotics, but all isolates were sensitive to vancomycin. All of MRSA had enterotoxin A in 30.6%, B 4.1%, C 8.3%, D, C/G, A 2.0% and None 51%. PFGE of SmaI-digested chromosomal DNA was performed on 49 sporadic MRSA isolates. Restriction fragment patterns consisted of 8 to 14 fragments ranged in size from 48.5 to 630.5 kbp. We could divided the isolates into 7 groups (<TEX>$I{\sim}VII$</TEX>) by analyzing PFGE patterns. Group I subdivided as 2 subgroups and 17 (34.7%) strains belong to the group I. Dendrogram of PFGE patterns showed that MRSA strains in Busan were heterogeneous but we could find out minor homogeneity in hospital.

In Vitro Activities of Ceftobiprole, Tigecycline, Daptomycin, and 19 Other Antimicrobials against Methicillin-Resistant Staphylococcus aureus Strains from a National Survey of Belgian Hospitals

The in vitro activities of 22 antimicrobial agents, including ceftobiprole, daptomycin, and tigecycline, against 511 methicillin-resistant Staphylococcus aureus (MRSA) isolates from 112 Belgian hospitals were studied by using the CLSI agar dilution method. Isolates were characterized by pulsed-field gel electrophoresis (PFGE) analysis and by PCR detection of determinants of resistance to aminoglycosides, macrolides-lincosamides-streptogramins, and tetracyclines. A representative set of isolates with different PFGE genotypes was further characterized by multilocus sequence typing, determination of staphylococcal cassette chromosome mec (SCCmec) type, and multiplex PCR for toxic shock syndrome type 1 (TSST-1) and Panton-Valentine leukocidin genes. MRSA isolates belonged to nine epidemic MRSA clones, of which sequence type 45 (ST45)-SCCmec IV and ST8-SCCmec IV were predominant, accounting for 49 and 20% of isolates, respectively. The distribution of antimicrobial resistance and TSST-1 genes was strongly linked to clonal types. Ceftobiprole, daptomycin, and tigecycline showed high activity against all isolates of these sporadic and epidemic MRSA clones, as indicated by MIC(90)s of 2 mg/liter, 0.5 mg/liter, and 0.25 mg/liter, respectively. The MIC distribution of daptomycin and tigecycline was not different in isolates with decreased susceptibility to glycopeptides or tetracyclines, respectively. Ceftobiprole MICs were not correlated with oxacillin and cefoxitin MICs. These data indicate excellent activity of the newly developed agents ceftobiprole, daptomycin, and tigecycline against MRSA isolates recently recovered from hospitalized patients in Belgium, supporting their therapeutic potential for nosocomial MRSA infections.

Evolution of sporadic isolates of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals and their similarities to isolates of community-acquired MRSA.

Forty-one methicillin-resistant Staphylococcus aureus (MRSA) hospital isolates that clearly differed from the six major pandemic clones of MRSA in pulsed-field gel electrophoresis type, mecA and Tn554 polymorphism, and epidemic behavior were selected from an international strain collection for more detailed characterization. SpaA typing, multilocus sequence typing, and SCCmec (staphylococcal cassette chromosome mec) typing demonstrated extensive diversity among these sporadic isolates both in genetic background and also in the structure of the associated SCCmec elements. Nevertheless, the isolates could be grouped into restricted clonal complexes by using the BURST (i.e., based upon related sequence types) program algorithm, which predicted that most sporadic MRSA isolates evolved from pandemic MRSA clones. Several of the sporadic MRSA resembled community-acquired MRSA isolates in properties that included a relatively limited multiresistance pattern, faster growth rates, diversity of genetic backgrounds, and a frequent association with SCCmec type IV.

Clonal and antibiotic resistance profiles of methicillin-resistant Staphylococcus aureus (MRSA) from a Portuguese hospital over time.

Methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered from a general hospital in Oporto, Portugal, during two periods (1992-1993 and 1996-2000) were characterized by pulsed-field gel electrophoresis (PFGE) of SmaI fragments, and by hybridization of ClaI digests with mecA and Tn554 probes, discriminating the isolates in mecA::Tn554::PFGE genotypes. In addition, a representative sample of the defined genotypes was characterized by multilocus sequence typing (MLST) and SCCmec (staphylococcal cassette chromosome mec) typing, generating the corresponding ST-SCCmec types. In 1992-1993, 77% of MRSA belonged to the Iberian clone (genotype I::E::A or ST247-IA). In 1996-2000, the frequency of this clone decreased to 19% and the majority (69%) of the isolates belonged to another international clone, the Brazilian MRSA (genotype XI::B::B or ST239-IIIA). Trimethoprim/sulfamethoxazole (SXT) was confirmed to be an important phenotypic marker to distinguish the Iberian (SXT-susceptible) and the Brazilian (SXT-resistant) clones in MRSA isolates from Portugal. Our observations document major shifts in the dominant MRSA clonal types that occurred in this hospital since 1992, suggesting a selective advantage of the Brazilian relatively to the Iberian clone. In addition to these two MRSA clones that are the most frequent in Portuguese hospitals since the early 1990s, sporadic MRSA clones (representing 14% of the total) were identified and characterized.

Better environmental survival of outbreak vs. sporadic MRSA isolates

Environmental sources have been associated with prolonged epidemics of methicillin-resistantStaphylococcus aureus (MRSA). Our objective was to evaluate whether outbreak MRSA strains differ in their environmental survival from sporadic MRSA strains. The strains were detected in five adult patients unknowingly colonized by MRSA. The MRSA strains from patients No. 1 (phage pattern; III-29) and No. 2 (III-215) caused extensive outbreaks in our hospital. Contact tracing after detection of the different MRSA strains in the three other patients did not reveal further spread.Suspensions of MRSA with and without added hospital dust were made in sterile PBS. Surviving counts of MRSA were made by culture at two-weekly or longer intervals up to approximately one year. For each MRSA strain the ultimate day of viability was ascertained. The environmental survival patterns of the five MRSA strains showed qualitative and quantitative differences between the two outbreak and three sporadic strains. A gradual decline was noted for all strains. All survived longer than 6 months, but the two outbreak strains survived significantly better (P<0.01), i.e. in higher quantities (circa 1000-fold) and for a 1–3 months longer period. The survival patterns of the MRSA strains with and without added dust were similar.

Molecular analysis of the accessory gene regulator (agr) locus and balance of virulence factor expression in epidemic methicillin-resistant Staphylococcus aureus.

Potential relationships between virulence factor expression and transmissibility were assessed in epidemic methicillin-resistant Staphylococcus aureus (MRSA) clones CMRSA-1 and CMRSA-3. A major subtype of CMRSA-1 exhibited normal transcription of RNAIII, which facilitates the induction of secreted virulence factors and repression of colonization factor expression at high cell density. However, these isolates characteristically did not express alpha-toxin or protease and displayed a limited profile of secreted proteins. CMRSA-1 also expressed a novel cell surface glycoprotein and exhibited a unique polymorphism within the accessory gene regulator (agr) locus. CMRSA-3 displayed attenuated activation of RNAIII transcription, which was consistent with its higher fibronectin-binding and coagulase activity relative to sporadic MRSA or CMRSA-1 (P=.05), low protease activity, and limited profile of secreted proteins. Thus, the balance of virulence factor expression in CMRSA-1 and CMRSA-3 favors the colonization phase of infection, and CMRSA-1 possesses unique genotypic and phenotypic traits.