Capillary electrophoretic methods for classification of methicillin-resistant Staphylococcus aureus (MRSA) clones

Aims: This study evaluated the influence of exposure to the hospital environment on methicillin-resistant Staphylococcus aureus (MRSA) carriage. The antibiograms of the MRSA isolates were examined. Materials and Methods: Nasal, throat, and web-space swabs were collected from 119 nursing students of the age group 18-23 years (exposed group) and 100 age-matched pharmacy students (nonexposed group). S. aureus was identified and antibiogram obtained as per Clinical and Laboratory Standards Institute (CLSI) guidelines. MRSA was detected by cefoxitin disc diffusion test and by growth on oxacillin screen agar as per CLSI guidelines. The presence of the mecA gene was confirmed by conventional polymerase chain reaction. Results: The MRSA carrier rates were 11.8% and 4% in the exposed and nonexposed groups, respectively. Association of exposure to the hospital environment with MRSA colonization was statistically significant. All MRSA isolates showed sensitivity to netilmicin, linezolid, tetracycline, vancomycin and teicoplanin. Among the exposed group, 71.4% MRSA isolates were resistant to ciprofloxacin, 64.3% to cotrimoxazole, 64.3% to erythromycin, 28.6% to gentamicin and 21.4% to clindamycin. Among the nonexposed group, 75% MRSA isolates were resistant to ciprofloxacin, 25% to cotrimoxazole, 25% to erythromycin, 25% to gentamicin and 25% to clindamycin. Conclusion: Exposure to the hospital environment was found to be a significant risk factor for MRSA carriage. Hospital-acquired MRSA (HA-MRSA) isolates showed greater resistance toward antimicrobials compared with community-acquired MRSA (CA-MRSA) isolates. This highlights the need for the appropriate institution of pharmacotherapy in cases of HA-MRSA and CA-MRSA infections and control of transmission by carriers.

Background: Nosocomial infections caused by methicillin-resistant Staphylococci could lead to increased morbidity and mortality, but little is known about the prevalence of infections with these organisms in healthcare facilities and in the community in Tripoli. This study investigated the in vitro susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase negative staphylococci (MRCNS) to antimicrobial agents, and determined the molecular characteristics of MRSA. Methods: This is a retrospective observational study aiming at determining the prevalence and antibiotic resistance pattern of (MRSA) and (MRCNS) isolated from non-duplicated clinical specimens in Tripoli Central Hospital (TCH) between June 2013 and June 2014. Isolates were identified using standard laboratory procedures. Antimicrobial susceptibility tests were carried out by disk diffusion method and automated systems. DNA of the MRSA isolates was used for PCR to determine the molecular analysis. Results: 218 isolates of Staphylococci were obtained, 71.6% were coagulase positive staphylococci (CPS) and 28.4% were coagulase negative staphylococci (CNS). 39.7% of CPS were MRSA, while 75.8% of CNS were MRCNS. The rates of hospital-acquired MRSA (HA-MRSA) and community-acquired MRSA (CA-MRSA) among MRSA isolates were 61.3% and 38.7% respectively. A similar trend was detected among MRCNS isolates, where 74.5% were HA-MRCNS and 25.5% were CA-MRCNS. All the MRSA and MRCNS isolates were susceptible (100%) to vancomycin, tigecycline, linezolid, quinupristin/dalfopristin, daptomycin and moxifloxacin. Generally, hospital-acquired strains showed higher resistance rates than community-acquired ones to the most commonly tested non-beta-lactam antibiotics. 35.5% of all staphylococcal isolates exhibited mecA+ gene and 12.9% expressed mecC+. Meanwhile, 38.7% of MRSA isolates harbored both mecA and mecC. However, 12.9% of MSSA isolates were negative for both mecA and mecC. The mecA gene was detectable in 59.1% and 40.9 % of HA-MRSA and CA-MRSA isolates respectively. Conclusion: Hospital-acquired MRSA and MRCNS isolates had higher resistance rates to non-beta lactam antimicrobial drugs than the respective community-acquired isolates. This was shown by early detection of mecC gene among MRSA isolates.

Background: Methicillin Resistant Staphylococcus aureus (MRSA) has evolved as a serious threat to public health. Objectives: The objectives of the present study were to determine the antibiotics susceptibility patterns of S. aureus and the minimum inhibitory concentration (MIC) of vancomycin to MRSA isolated from different clinical samples at the Colombo South Teaching Hospital (CSTH) in Sri Lanka. Methodology: A total 72 isolates of S. aureus, obtained from different clinical samples at the CSTH, from January to May 2017 were included in the study. S. aureus isolates were identified by Gram stain, colony morphology, catalase, slide/tube coagulase tests. The antibiotic susceptibility tests were carried as per Clinical Laboratory Standards Institute (CLSI) guidelines. MRSA isolates were detected using the cefoxitin (30 μg) disk diffusion test. Inducible clindamycin resistance (MLSB i) was detected by the disk approximation (D test) test. The vancomycin MICs were determined by the E-test method with a 0.5 McFarland standard inoculum. The MIC clinical breakpoints were defined according to the CLSI guidelines (susceptible, ≤2 μg/ml; intermediate, 4–8 μg/ml; and resistant, ≥16 μg/ml). Results: Of the 72 S. aureus clinical isolates, 29 (40.2%) were MRSA. Inducible clindamycin resistance was detected in 16% of the MRSA isolates. Minimum inhibitory concentrations of vancomycin to the isolates of MRSA ranged from 0.125 μg/ml to 2 μg/ml. Conclusions: The rate of isolation of MRSA was high and it has emerged as a serious public health threat to Sri Lanka. The Minimum Inhibitory Concentration (MIC) of all the MRSA isolates were ≤2 μg/ml. None of the MRSA isolates were found to be intermediate-sensitive or vancomycin resistant. Therefore, vancomycin can be used as the drug of choice for treatment of infections caused by MRSA.

Background: Trimethoprim-sulfamethoxazole is commonly used for the treatment of noninvasive methicillin-resistant Staphylococcus aureus (MRSA) infections. Following a report from 2 facilities of increased trimethoprim-sulfamethoxazole resistance among MRSA infections, we assessed changes in resistance nationally and by state. Methods: We reviewed antibiotic susceptibility testing (AST) data for trimethoprim-sulfamethoxazole among S. aureus isolates associated with surgical site infections (SSIs), central-line–associated bloodstream infections (CLABSIs), and catheter-associated urinary tract infections (CAUTIs) from acute-care hospitals reported to the NHSN Device and Procedure Module from 2012 to 2018. We compared the pooled mean percentage of isolates nonsusceptible to trimethoprim-sulfamethoxazole in 2012 and 2018, stratified by MRSA and methicillin-sensitive Staphylococcus aureus (MSSA). Among MRSA isolates, we compared the percentage nonsusceptible to trimethoprim-sulfamethoxazole by healthcare-associated infection (HAI) type and state in 2012 and 2018. States with ≥20 MRSA isolates with AST reported each year were included in the state-level analysis. Results: Overall, 36,587 MRSA isolates and 46,824 MSSA isolates were reported from 2012 to 2018. Moreover, >80% of MRSA and MSSA isolates had trimethoprim-sulfamethoxazole AST reported each year. Nationally, the percentage of trimethoprim-sulfamethoxazole nonsusceptible among MRSA isolates was 3.9% in 2012 compared to 6.5% in 2018 (P < .001), but it was unchanged among MSSA isolates during the same period (1.1% in 2012 vs 1.4% in 2018; P = .08). Among MRSA surgical site infections (SSIs), the proportion of trimethoprim-sulfamethoxazole nonsusceptible isolates was 3.1% in 2012 versus 6.1% in 2018 (P < .001) but did not change significantly for CLABSIs or CAUTIs (Fig. 1). Among the 32 states that met the inclusion criteria, there were no significant decreases, whereas 4 (12.5%) showed significant increases in the percentage of MRSA that were trimethoprim-sulfamethoxazole nonsusceptible in 2018 compared to 2012: New Jersey (2.4% in 2012 vs 19.3% in 2018; P <.001); Florida (9.1% in 2012 vs 22.4% in 2018; P < .001); Maryland (0.0% in 2012 vs 10.9% in 2018; P < .01); and Pennsylvania (1.7% in 2012 vs 6.5% in 2018; P < .001). Conclusions: Nationally, there was a modest but significant increase in the percentage of MRSA HAI isolates nonsusceptible to trimethoprim-sulfamethoxazole in 2018 compared to 2012; however, 3 of 4 states with significant increases in nonsusceptibility had substantial, potentially clinically relevant increases (>10%). Ongoing characterization of MRSA isolates from Florida and New Jersey may provide insight into the underlying cause of these shifting patterns in trimethoprim-sulfamethoxazole resistance among MRSA. Healthcare personnel should select appropriate antibiotic regimens based on local resistance patterns, should monitor patients for treatment failure, and should report changes in resistance to the appropriate public health department.Funding: NoneDisclosures: None

PurposeTo evaluate the rate of nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) in healthy children and to determine the staphylococcal chromosomal cassette mec (SCCmec) type and toxin profile of these...

A comparison of previous antibiotic therapy following isolation of MRSA versus MSSA in nursing home residents: a preliminary investigation.

with Atypical Resistance Profi le, Spain

BackgroundDespite a substantial burden of non-bacteraemic methicillin resistant Staphylococcus aureus (MRSA) disease, most MRSA surveillance schemes are based on bacteraemias. Using bacteraemia as an outcome, trends at hospital level are difficult to discern, due to random variation. We investigated rates of nosocomial bacteraemic and non-bacteraemic MRSA infection as surveillance outcomes.Methods and FindingsWe used microbiology and patient administration system data from an Oxford hospital to estimate monthly rates of first nosocomial MRSA bacteraemia, and nosocomial MRSA isolation from blood/respiratory/sterile site specimens (“sterile sites”) or all clinical samples (screens excluded) in all patients admitted from the community for at least 2 days between April 1998 and June 2006. During this period there were 441 nosocomial MRSA bacteraemias, 1464 MRSA isolations from sterile sites, and 3450 isolations from clinical specimens (8% blood, 15% sterile site, 10% respiratory, 59% surface swabs, 8% urine) in over 2.6 million patient-days. The ratio of bacteraemias to sterile site and all clinical isolations was similar over this period (around 3 and 8-fold lower respectively), during which rates of nosocomial MRSA bacteraemia increased by 27% per year to July 2003 before decreasing by 18% per year thereafter (heterogeneity p<0.001). Trends in sterile site and all clinical isolations were similar. Notably, a change in rate of all clinical MRSA isolations in December 2002 could first be detected with conventional statistical significance by August 2003 (p = 0.03). In contrast, when monitoring MRSA bacteraemia, identification of probable changes in trend took longer, first achieving p<0.05 in July 2004.ConclusionsMRSA isolation from all sites of suspected infection, including bacteraemic and non-bacteraemic isolation, is a potential new surveillance method for MRSA control. It occurs about 8 times more frequently than bacteraemia, allowing robust statistical determination of changing rates over substantially shorter times or smaller areas than using bacteraemia as an outcome.

BackgroundMethicillin resistant Staphylococcus aureus (MRSA) is a major human pathogen associated with nosocomial and community infections. Panton Valentine leukocidin (PVL) is considered one of the important virulence factors of S. aureus responsible for destruction of white blood cells, necrosis and apoptosis and as a marker of community acquired MRSA. This study was aimed to determine the prevalence of PVL genes among MRSA isolates and to check the reliability of PVL as marker of community acquired MRSA isolates from Western Nepal.MethodsA total of 400 strains of S. aureus were collected from clinical specimens and various units (Operation Theater, Intensive Care Units) of the hospital and 139 of these had been confirmed as MRSA by previous study. Multiplex PCR was used to detect mecA and PVL genes. Clinical data as well as antimicrobial susceptibility data was analyzed and compared among PVL positive and negative MRSA isolates.ResultsOut of 139 MRSA isolates, 79 (56.8 %) were PVL positive. The majority of the community acquired MRSA (90.4 %) were PVL positive (Positive predictive value: 94.9 % and negative predictive value: 86.6 %), while PVL was detected only in 4 (7.1 %) hospital associated MRSA strains. None of the MRSA isolates from hospital environment was found positive for the PVL genes. The majority of the PVL positive strains (75.5 %) were isolated from pus samples. Antibiotic resistance among PVL negative MRSA isolates was found higher as compared to PVL positive MRSA.ConclusionOur study showed high prevalence of PVL among community acquired MRSA isolates. Absence of PVL among MRSA isolates from hospital environment indicates its poor association with hospital acquired MRSA and therefore, PVL may be used a marker for community acquired MRSA. This is first study from Nepal, to test PVL among MRSA isolates from hospital environment.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-016-1531-1) contains supplementary material, which is available to authorized users.

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The increase in the prevalence of epidemic strains of methicillin resistant Staphylococcus aureus (MRSA) in hospitals and community requires special attention of infection control. The aim of this study was to determine the pathogenic phenotype (i.e. infectivity and resistotype) and genotypic characteristics (i.e. PFGE-pulsotyping, SLST-spa typing, MLST-sequence typing, eBURST-clonal complex detection algorithm) of clinical MRSA isolates in the Central Blacksea region of Turkey, in order to understand their short- and long-term epidemiological and evolutionary dynamics, and to investigate any probable presence of a significant clustering. This prospective study included consecutive but non-repetitive 48 MRSA isolates (of them 18 were colonized strains and 30 were causes of nosocomial infection) and seven methicillin-susceptible S.aureus (MSSA, all were isolated from nosocomial infection), collected between December 2006-February 2007 period from hospitalized patients. Identification of the isolates were performed by Vitek-2 automated system (BioMérieux, USA), and in vitro antimicrobial susceptibility testing by broth microdilution method and Vitek-2 automated system. The MRSA isolates found susceptible to erythromycin (n= 10) were further investigated for the presence of ermA gene by the PCR method. All the strains were typed by spa-typing and PFGE-pulsotyping methods. Among the isolates with different spa-types, representatives were selected (3 MRSA, 7 MSSA) and typed with MLST typing method. Among the isolates with different spa-types, representatives with different antimicrobial susceptibility patterns were selected (n= 8), and SCCmec types were determined by the multiplex PCR method. Antimicrobial resistance patterns of the isolates were digitized to get standardized antimicrobial resistance phenotypes. Clustering of MRSA isolates in pattern groups on the basis of discriminatory characteristics, namely infectivity, phenotype and genotype were statistically analyzed with specific inclusion and exclusion criteria. As a result, three different antimicrobial resistance phenotypes were found in MSSA isolates, whereas 13 were identified in MRSA isolates. In MSSA isolates, seven different PFGE-pulsotypes were detected, as compared to 14 pulsotypes in MRSA isolates. Among MRSA isolates, 10 sporadic strains with single PFGE-pulsotypes were detected. All MRSA isolates, with two exceptions (t459, t632), were of t030 spa-type; in the MLST analysis of the representatives of different spa-types (n= 3), a single type of MLST-clonal complex (CC8) and single MLST-sequence type (ST239) were identified. Each of the seven MSSA isolates yielded different spa-types, MLST-clonal complex types and MLST-sequence types (t777-ST5-CC5; t660-ST25-CC5; t153-ST34-CC30; t015-ST45-CC45; t267-ST97-CC97; t377-ST360-CC8; t084-ST15-C15). In the statistical analysis of 38 non-sporadic MRSA isolates, the isolates in Group-13 (n= 16; infectious, resistotype 14, pulsotype 4; antimicrobial resistance score= 24) displayed significant infectivity-phenotype-genotype clustering (p< 0.001). In 27 of the MRSA isolates, decreased susceptibility to teicoplanin (MIC= 4 µg/mL) was detected. Although, global MRSA isolates belonging to MLST-CC8, MLST-ST239, t030 spa-type were usually expected to be resistant to erythromycin, 10 such strains were erythromycin susceptible. However, ermA gene was found in six of these 10 strains, leading to a conclusion that the ermA gene of these isolates might be dysfunctional due to a point mutation or deletion. Selected representatives of MRSA isolates with different antimicrobial susceptibility patterns (n= 8) were detected to be SCCmec type III. In conclusion, S.aureus isolates in the patient population of our hospital representing the Central Blacksea region showed statistically significant clustering in infectivity, antimicrobial resistance phenotype and clonal genotype (p< 0.001). The dominant MRSA clone was ST239 which was one of the five major pandemic MRSA clones. Nosocomial MSSA isolates displayed long-term clonal diversity. This study produced regional evolutionary-epidemiological data that may support further regional, national and international long-term surveillance studies of S.aureus strains.

To identify the staphylococcal cassette chromosome mec (SCCmec) types of methicillin-resistant Staphylococcus aureus (MRSA) isolated from bovine milk, and examine the genetic relatedness between MRSA from bovine milk and MRSA from human isolates. Antimicrobial susceptibility tests were performed on MRSA isolated from bovine milk. PCR and sequencing analysis were performed to determine the SCCmec type of MRSA, and to confirm their toxin carriage. Genetic relatedness among the bovine isolates and between bovine and human isolates was detected with PFGE and multilocus sequence typing (MLST). Fourteen MRSA and a silent mecA-carrying methicillin-susceptible S. aureus (smMSSA) were isolated from the milk of cows with an isolation ratio of 0.18%. SCCmec of 14 MRSA strains were designated as new subtype IVg, and one smMSSA strain was not classified. All 14 MRSA strains shared Panton-Valentine leucocidin (PVL) and staphylococcal enterotoxin D (SED), SEI and SEJ; the smMSSA strain had only PVL. All MRSA and smMSSA isolates showed no multidrug resistance and had community-acquired MRSA (CA-MRSA) characteristics. PFGE revealed that all isolates except the smMSSA belonged to the same genetic lineage, and MLST analysis showed that they had no genetic relatedness with CA-MRSA which had caused human infection in Korea. MRSA isolated from bovine milk harboured a unique SCCmec subtype, and they may not be correlated with the emergence of CA-MRSA in human infection in Korea.

Vancomycin Heteroresistance in Methicillin-resistant Staphylococcus aureus, Taiwan

In methicillin-resistant Staphylococcus aureus (MRSA) treatment, the vancomycin minimum inhibitory concentration (MIC) increase, vancomycin heteroresistance (hVISA) presence, and accessory gene regulator (agr) dysfunction are predictors of vancomycin therapy failure. This study evaluated the association between vancomycin MIC (≥ 1.0μg/mL) and agr dysfunction in invasive MRSA isolates. Vancomycin MIC, hVISA phenotype, agr group, and function were determined in 171 MRSA isolates obtained between 2014 and 2019 from hospitals in Porto Alegre, Brazil. All MRSA were susceptible to vancomycin; 16.4% of these had MIC ≥ 1.0μg/mL. Seventeen MRSA isolates expressed the hVISA phenotype; 35.3% of them had MIC of 1.5μg/mL. agr groups I (40.9%) and II (47.1%) were the most found groups for MRSA and hVISA isolates, respectively. The proportion of MRSA with vancomycin MIC ≥ 1.0μg/mL in agr group II was significantly higher than in agr groups I and III (p = 0.002). agr dysfunction was observed in 4.7% (8/171) of MRSA, especially those with vancomycin MIC ≥ 1.0μg/mL (p < 0.001). In addition, six isolates (35.3%; 6/17) with hVISA phenotype presented agr dysfunction, which was significantly higher than that in non-hVISA phenotype (p < 0.001). In conclusion, agr dysfunction in MRSA is associated with vancomycin MIC ≥ 1.0μg/mL and hVISA phenotype, which suggests that agr dysfunction might confer potential advantages on MRSA to survive in invasive infections.

Complex Molecular Epidemiology of Methicillin-Resistant Staphylococcus aureus Isolates From Children With Cystic Fibrosis in the Era of Epidemic Community-Associated Methicillin-Resistant S aureus