PVL-positive Methicillin-resistant Staphylococcus Aureus Research Articles

Significant increase in the prevalence of Panton–Valentine leukocidin-positive methicillin-resistant Staphylococcus aureus , particularly the USA300 variant ΨUSA300, in the Japanese community

ABSTRACT USA300, a major Panton–Valentine leukocidin (PVL)-producing methicillin-resistant Staphylococcus aureus (MRSA) clone associated with skin and soft tissue infections (SSTIs), carries a type IV staphylococcal cassette chromosome mec (SCC mec ) element. Recently, a USA300 variant, ΨUSA300, with a 12-bp deletion of ccrB2 in the SCC mec element, was identified in Japan by Takadama et al. (15). To understand the current status of PVL-positive MRSA in the community, a nationwide surveillance of MRSA isolated from outpatients at dermatology departments was conducted. This study included 1,619 outpatients who visited dermatology departments at 22 healthcare facilities in 11 prefectures in Japan between 2018 and 2021; S. aureus were isolated from these outpatients. Molecular epidemiological analysis was performed using PCR, SCC mec and spa typing, pulsed-field gel electrophoresis, multilocus sequence typing, and antimicrobial susceptibility testing. A total of 980 S . aureus were isolated, 293 of which were identified as MRSA. Among MRSA isolates, PVL genes were detected in 18 of 63 (28.6%) in 2018, 34 of 85 (40.0%) in 2019, 29 of 65 (44.6%) in 2020, and 40 in 80 (50.0%) in 2021. Among the PVL-positive MRSA, 16 of 121 (13.2%) and 94 of 121 (77.7%) isolates were identified as USA300 and ΨUSA300, respectively. ΨUSA300 strains were collected from six prefectures in Japan: Kagawa, Osaka, Tokyo, Kanagawa, Fukushima, and Saitama. ΨUSA300 was isolated more frequently than USA300 from patients with deep-seated SSTIs ( P < 0.05). Our findings showed that PVL-positive MRSA isolates were significantly increased in the Japanese community. Furthermore, the USA300 variant ΨUSA300 has emerged as the predominant PVL-positive MRSA clone across the Kanto, Kinki, and Shikoku regions of Japan, and this variant may possess a greater propensity to cause deep-seated SSTIs than in USA300. IMPORTANCE USA300 is an MRSA clone producing PVL, a toxin associated with SSTIs. ΨUSA300 is a USA300 variant recently identified in Japan by Takadama et al. (15). Here, we found that the prevalence rate of PVL-positive MRSA in S. aureus was elevated in the Japanese community, and ΨUSA300 accounted for most of them. ΨUSA300 strains have been isolated from several areas in Japan and were associated with deep-seated SSTIs. This study highlighted the emerging threat posed by ΨUSA300 in Japan.

Molecular characterization of MRSA collected during national surveillance between 2008 and 2019 in the Netherlands

Background.Although the Netherlands is a country with a low endemic level, methicillin-resistant Staphylococcus aureus (MRSA) poses a significant health care problem. Therefore, high coverage national MRSA surveillance has been in place since 1989. To monitor possible changes in the type-distribution and emergence of resistance and virulence, MRSA isolates are molecularly characterized.Methods.All 43,321 isolates from 36,520 persons, collected 2008–2019, were typed by multiple-locus variable number tandem repeats analysis (MLVA) with simultaneous PCR detection of the mecA, mecC and lukF-PV genes, indicative for PVL. Next-generation sequencing data of 4991 isolates from 4798 persons were used for whole genome multi-locus sequence typing (wgMLST) and identification of resistance and virulence genes.Results.We show temporal change in the molecular characteristics of the MRSA population with the proportion of PVL-positive isolates increasing from 15% in 2008–2010 to 25% in 2017–2019. In livestock-associated MRSA obtained from humans, PVL-positivity increases to 6% in 2017–2019 with isolates predominantly from regions with few pig farms. wgMLST reveals the presence of 35 genogroups with distinct resistance, virulence gene profiles and specimen origin. Typing shows prolonged persistent MRSA carriage with a mean carriage period of 407 days. There is a clear spatial and a weak temporal relationship between isolates that clustered in wgMLST, indicative for regional spread of MRSA strains.Conclusions.Using molecular characterization, this exceptionally large study shows genomic changes in the MRSA population at the national level. It reveals waxing and waning of types and genogroups and an increasing proportion of PVL-positive MRSA.

Mandatory Notification of Panton-Valentine Leukocidin-Positive Methicillin-Resistant Staphylococcus aureus in Saxony, Germany: Analysis of Cases from the City of Leipzig in 2019.

In Germany, Saxony is the only federal state where the detection of a Panton-Valentine Leukocidin (PVL)-positive Methicillin-resistant Staphylococcus aureus (MRSA) has to be notified to the local health authority (LHA). The LHA reports the case to the state health authority and introduces concrete infection control measures. We analyzed isolates from the respective cases in 2019, which were collected in local microbiological laboratories and sent to the National Reference Centre (NRC) for Staphylococci and Enterococci for strain characterization and typing. Antibiotic resistance testing was done by broth microdilution. Molecular characterization was performed using spa and SCCmec typing, MLST, and the PCR detection of marker genes associated with distinct MRSA lineages. Demographic and clinical data of the individual cases were assessed and the LHA performed epidemiological investigations. Thirty-nine (index) persons, diagnosed with a PVL-positive MRSA, were initially reported to the LHA. Most patients suffered from skin and soft-tissue infections (SSTI). For 21 of the index cases, household contacts were screened for MRSA. Seventeen out of 62 contacts were also colonized with a PVL-positive MRSA. The median age of altogether 58 individuals was 23.5 years. In over half of the cases, the home country was not Germany and/or a history of travel or migration was reported. Molecular characterization revealed the presence of various epidemic community-associated MRSA lineages, with "USA300", including the North American Epidemic (ST8-MRSA-IVa) and the South American Epidemic Clone (ST8-MRSA-IVc), the "Sri Lankan Clone" (ST5-MRSA-IVc), and the "Bengal Bay Clone" (ST772-MRSA-V) being more prevalent. In eight out of nine households, the contact persons were colonized with the same clone as the respective index case, suggesting a close epidemic and microbiological link. The obligation to report PVL-positive MRSA enables us to detect the occurrence of PVL-producing MRSA and its spread in the population as early as possible. Timely detection allows the targeted deployment of reliable anti-infective measures.

Predominance of t355/ST152/SCCmec V clonal type among PVL-positive MRSA isolates in a tertiary care hospital in Belgrade, Serbia.

Epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) is continually changing. Frequency of genotypes typical for community-associated MRSA (CA-MRSA) is increasing in hospitals, as well as resistance to antimicrobial agents. Moreover, different clones predominate in different geographic regions, and temporal shifts occur in the predominant clonal type. The aim of this study was to estimate the prevalence of MRSA, CA-MRSA and PVL-positive MRSA isolates from patients hospitalised in the Military Medical Academy (MMA) and from outpatients, and to perform genotyping of PVL-positive MRSA isolates. MRSA isolates were obtained by standard microbiological techniques. PVL-positive MRSA were detected by single PCR. Determination of SCCmec types in MRSA isolates was done using multiplex PCR and genotyping of PVL-positive MRSA by PFGE, MLST and spa typing. The prevalence of MRSA among S. aureus isolates from different clinical specimens was 43.4%. In outpatients the prevalence of MRSA was 3.2%. SCCmec types specific for CA-MRSA were found in 26% of MRSA isolates from hospitalised patients. In groups, hospitalised patients and outpatients, the prevalence of PVL-positive MRSA isolates was 4%, and all of them harboured SCCmec type V genetic element. PFGE revealed minor differences between four groups of PVL-positive MRSA isolates, but all of them belonged to ST152, and all except one were of the t355 spa type. High prevalence of MRSA and CA-MRSA in MMA, especially the presence of PVL-positive CA-MRSA, represent a serious health threat for patients. Genotype t355/ST152/SCCmec V is the dominant MRSA clone among PVL-positive CA-MRSA.

Clinical features and antimicrobial treatment of skin infections caused by Panton‐Valentine leukocidin‐positive methicillin‐resistant Staphylococcus aureus

Skin infections caused by Panton-Valentine leukocidin (PVL)-positive methicillin-resistant Staphylococcus aureus (MRSA), especially the USA300 clone, have been increasing in Japan. To prevent an epidemic of PVL-positive MRSA, rapid diagnosis and effective antimicrobial therapy are essential. However, the clinical features of, and antimicrobial efficacy against, these skin infections are not well understood in Japan. Here, we report 10 cases of skin infections caused by PVL-positive MRSA that presented over a two-year period in our clinic. Genetic analyses revealed that 90% of the PVL-positive MRSA strains were identified as USA300 and its related clones. Notably, 70% of the patients had atopic dermatitis (AD) as an underlying disease. Average durations of antimicrobial therapy for AD patients (10.6 weeks) were 2.9-fold longer than those for non-AD patients (3.7 weeks). However, all cases were improved by a long-term course of fosfomycin, minocycline, doxycycline, and/or rifampicin. Our data suggest that AD may be an important risk factor for intractable skin infections caused by PVL-positive MRSA.

Epidemiology of the Staphylococcus aureus CA-MRSA USA300 in Belgium.

The methicillin-resistant Staphylococcus aureus (MRSA) sequence type (ST) 8 Panton-Valentine toxin (PVL)-positive USA300 clone has a worldwide distribution. The USA300 North American (NA) variant, harbouring the arginine catabolic mobile element (ACME), is predominant in the USA while the Latin American (LV) variant is predominant in Northern South America. Both variants have failed to become endemic in Europe. We examined here the epidemiology of the USA300 clone in Belgium from 2006 to 2019. A total of 399 clonal complex 8 PVL-positive MRSA isolates received between 2006 and 2019 by the Belgian National Reference Laboratory for S. aureus were investigated for the presence of ACME. Selected ACME-positive (n=102) and ACME-negative (n=16) isolates were sequenced, characterized for the presence of several resistance and virulence molecular markers and subjected to phylogenetic analysis. A total of 300 isolates were USA300-NA (ACME-positive), while only 99 were ACME-negative. Most USA300-NA interspersed in the phylogeny analysis with isolates from other countries, suggesting multiple introductions. However, two big clades were maintained and spread over a decade, peaking between 2010 and 2017 to finally decrease. Few ACME-negative isolates, mainly related to trips to South America, were identified as USA300-LV. The remaining ACME-negative isolates were ST8 SCCmec IVb or ST923 SCCmec IVa (COL923). Two clades of the USA300-NA clone have successfully spread in Belgium, but seem to currently decrease. Related South American variants have been detected for the first time in Belgium, including the emerging COL923 clone.

Colonization and Genetic Diversity of MRSA Among ICU Patients and Healthcare Workers From a Hospital of Northeastern India

Background: The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) is diverse in different geographic regions, and transmission of MRSA in hospital settings occurs either through the patients or asymptomatic healthcare workers who MRSA colonized in the nares and thus act as a potential reservoir of infection for susceptible patients. Methods:S. aureus isolates were collected from ICU patients and healthcare workers at Gauhati Medical College and Hospital, Assam, India, from May 2010 to April 2015. Premoistened swab samples were obtained from the ICU patients from 4 different sites (ie, nares, axilla, groin, and perianal region) within 48 hours of admission. For healthcare workers (HCWs) a nasal swab was obtained. The isolates were identified by phenotypic and genotypic methods using CLSI guidelines and PCR (fem B, MecA, and PVL). The antibiograms were obtained using a Vitek 2 system. Results: For 84 patients admitted to the ICU, swab samples were obtained from various sites, and S. aureus was observed in 34 samples (40.5%). Among the isolates, 13 (38%) were MRSA and 21 (62%) were methicillin-susceptible Staphylococcus aureus (MSSA). Among the HCWs from the ICU, growth of S. aureus was obtained in 10 of 30 samples (33.34%), of which 3 (30%) were MRSA and 7 (70%) were MSSA. S. aureus isolates were genotypically identified as fem B among colonized patients (40.5%) and HCWs (33.34%). MRSA (mecA positive) was detected in 3% of colonized patients and 30% of HCWs. Among the ICU patients, 78.56% were PVL-positive S. aureus: 21.42% were PVL-positive MRSA and 57.14% were PVL-positive MSSA. Multilocus sequence typing of the 7 housekeeping genes against 2 S. aureus isolates showed the presence of ST1428, which had not been reported in India, whereas the other sequence was entirely novel. The MDR rates were 68% and 75% among ICU patients and HCWs, respectively, and all the strains were mupirocin sensitive. The S. aureus isolates were significantly proportional among HCWs compared to the colonized group (P = .031). Conclusions: The study results show a high prevalence of PVL-positive MSSA and MRSA among ICU patients. This finding indicates its transmission among hospitalized patients through the HCWs, for which constant monitoring of the pathogen, particularly its phenotypic and genotypic variations and antimicrobial resistance pattern, is needed to develop effective strategies for infection prevention.Funding: NoneDisclosures: None

Prevalence and association of Panton-Valentine Leukocidin gene with the risk of sepsis in patients infected with Methicillin Resistant Staphylococcus aureus

BackgroundPanton-Valentine Leukocidin (PVL), is one of the virulence gene expressed by Methicillin Resistant Staphylococcus aureus (MRSA) and is known to be associated with severe form of community acquired MRSA infection. The aim of this study is to investigate its prevalence in our setting and patient’s clinical outcome. MethodsA cross sectional study involve retrospective record review were done involving 90 MRSA positive isolates between November 2016 and October 2017. Multiplex PCR was performed to detect femA, mecA and PVL genes. Clinical presentation and outcomes of patients were reviewed and presented as descriptive analysis. ResultsAll of the 90 MRSA isolates included in this study were positive for femA and mecA genes following PCR. PVL gene was detected in 20% (n = 18) of the isolates of which 61.1% (n = 11) were community acquired infections and 38.8% (n = 7) were hospital acquired. Case distribution from community acquired infections include patients with skin and soft tissue infections (33.3%, n = 6), infected diabetic foot ulcers (16.7%, n = 3), and one patient each (5.5%, n = 1) for community acquired pneumonia and meningitis. Half of the PVL positive MRSA cases (50%, n = 9) were having sepsis and four of them succumbed to death due to severe infection. ConclusionThis study shows a high prevalence of PVL positive MRSA infection in our population. Skin and soft tissue infections accounting for the major sources. In addition, the presence of the PVL gene is associated with increased risk for developing sepsis.

Draft Genome Sequence of the Panton-Valentine Leucocidin-Producing Staphylococcus aureus Sequence Type 154 Strain NRL 08/001, Isolated from a Fatal Case of Necrotizing Pneumonia.

Panton-Valentine leucocidin (PVL)-positive methicillin-resistant Staphylococcus aureus (MRSA) strains cause life-threatening diseases. We present a draft genome sequence of PVL-positive MRSA sequence type 154 (ST154) strain NRL 08/001, isolated from a fatal case of necrotizing pneumonia. The genome consists of 2.9 Mb over 39 contigs and harbors novel composite island staphylococcal cassette chromosome mec element (SCCmec)-mercury composite type 2B&5.

MOLECULAR CHARACTERIZATION AND ANTIMICROBIAL SUSCEPTIBILITY OF MRSA ISOLATED FROM CHRONIC HEMODIALYSIS OUTPATIENTS AND THEIR CORRELATION TO MRSA COLONIZATION AMONG HEALTHCARE WORKERS

The carriage of methicillin-resistant Staphylococcus aureus (MRSA) among dialysis patients is remarkable not only in terms of the risks of developing infections, but also in playing a principle part in transmission among dialysis unit staff. The aim of this study was to detect the colonization of Methicillin-sensitive Staphylococcus aureus and MRSA carriage. Also, our aim was to determine the relatedness of MRSA isolates and the potential routes of transmission using PCR- Restriction Fragment Length Polymorphism (PCR-RFLP) in Hemodialysis Unit of El Zagazig General Hospital, a tertiary medical center in Sharqia, Egypt. This study was conducted on 150 chronic hemodialysis outpatients and 200 non clinical control samples including environmental and healthcare workers (HCWs). Antibiotic susceptibility by VITEK-2 and disc diffusion, PCR amplification of mecA, pvl and coa genes and RFLP-PCR were conducted during the study period. In this study 3.3% of the patients and 3.2% of HCWs colonized with pvl positive MRSA. Fifty percent of MRSA isolates showed a single band PCR product amplification of 810bp fragment corresponding to coa gene. Ten distinct MRSA RFLP banding patterns designated as H1-H10 were obtained. The majority of strains belonged to RFLP banding pattern H1 (33.33%).The prevalence of MRSA carriage among hemodialysis patients was 14% and 9.7 % among HCWs with similar polymorphism patterns. The presence of one major coa gene type confirmed the occurrence of hospital acquired-associated MRSA.

Increase in the prevalence of Panton–Valentine leukocidin and clonal shift in community-onset methicillin-resistant Staphylococcus aureus causing skin and soft-tissue infections in the Rhine-Neckar Region, Germany, 2012–2016

Methicillin-resistant Staphylococcus aureus (MRSA) remains a major challenge for patient care. Community-associated (CA)-MRSA often have a fitness and virulence advantage compared with their nosocomial counterparts. Increased mobility, travel activities and migration accelerate the intercontinental spread of virulent CA-MRSA strains. Outpatient clinics are the most important route of entry for CA-MRSA into hospitals. However, systematic data on CA-MRSA in Germany are limited. In this study, community-onset (CO)-MRSA skin and soft-tissue infection (SSTI) isolates in the Rhine-Neckar Region from 2012-2016 were characterised to gain an insight into their molecular epidemiology and to monitor potential introduction of virulent and dominant MRSA strains into our hospital. A total of 2475 patients with S. aureus SSTI were identified in the outpatient departments of our hospital, of which 94 (3.8%) were MRSA. In addition, 40.4% of the CO-MRSA harboured the virulence factor Panton-Valentine leukocidin (PVL). ST8-t008-MRSA-IVa/c (23.7%; 9/39) and ST80-t044-MRSA-IVc (15.8%; 6/38) were the predominant PVL-positive MRSA. Molecular typing and epidemiological data revealed that 42.6% (40/94) of strains could be traced back to a local origin and 44.7% (42/94) were endemic outside of Europe. Resistance to quinolones, clindamycin and macrolides was common, whilst resistance to trimethoprim/sulfamethoxazole, tetracycline, mupirocin, chlorhexidine and fusidic acid was low. No resistance to rifampicin, fosfomycin or linezolid was observed. This study provides insight into the clonal composition of CO-MRSA in the Rhine-Neckar Region. The increase of PVL-positive MRSA and the introduction of imported strains may affect the local MRSA landscape in the near future and should be monitored closely.

Dissemination of Panton-Valentine leukocidin–positive methicillin-resistant Staphylococcus aureus USA300 clone in multiple hospitals in Tokyo, Japan

ObjectivesPanton-Valentine leukocidin (PVL) gene (lukS/F-PV)-positive community-acquired methicillin-resistant Staphylococcus aureus (MRSA), particularly the USA300 clone, is a health concern worldwide. Recently, community-acquired MRSA clones have been found to spread and persist in hospital settings. We aimed to investigate the prevalence and molecular epidemiologic features of lukS/F-PV–positive MRSA in Japanese hospitals. MethodsA total of 3433 MRSA isolated from nine hospitals in 2011–2015 were assessed. Molecular epidemiologic analysis performed by staphylococcal cassette chromosome mec typing, PVL typing, arginine catabolic mobile element typing, detection of virulence determinants, multilocus sequence typing, pulsed-field gel electrophoresis and antimicrobial susceptibility testing. ResultsThe detection rate of lukS/F-PV was increased annually from 0.6% (5/817) in 2011 to 3.1% (17/544) in 2015. Molecular epidemiologic analysis for 64 lukS/F-PV–positive MRSA isolates revealed that 42 isolates (65.6%) were the USA300 clone. Resistance rates of levofloxacin and gentamicin among lukS/F-PV–positive isolates increased annually as a result of increased prevalence of the multidrug-resistant USA300 clone. ConclusionsThis is the first report on dissemination of the USA300 clone and PVL-positive MRSA in multiple Japanese hospitals. Our data strongly suggest that the USA300 clone may become epidemic in Japanese hospitals.

Molecular characterization of Panton-Valentine leukocidin (PVL) toxin-encoding phages from South India.

A total of 19 methicillin-resistant Staphylococcus aureus (MRSA) isolates were investigated for Panton-Valentine leukocidin (PVL) toxin, PVL gene sequence variation and PVL-encoding phages. Whole genome sequencing was performed for all isolates. Analysis of MRSA isolates (n = 19) confirmed that most MRSA (n = 11) were positive for the PVL gene and were multidrug resistant. ST772-MRSA-V was the predominant PVL-positive MRSA clone, although all of them were found to carry the ΦIND772PVL phage in the genome. This study provides insights into the evolution of a new lineage of PVL-MRSA and highlights the potential risk of the emergence of multidrug-resistant community-acquired MRSA with high virulence.

Epidemiology and molecular characteristics of methicillin-resistantStaphylococcus aureus from skin and soft tissue infections in Shiraz, Iran.

Panton-Valentine leukocidin (PVL)-positive methicillin-resistant Staphylococcus aureus (MRSA) infections are increasing in some regions of Iran. The aim of the current study was to assess the epidemiology and molecular characteristics of S. aureus isolated from patients with skin infections in Shiraz, Iran. Swab samples were obtained from patients admitted to the skin and burn units of hospitals. The medical records of each patient were collected via questionnaire. All staphylococcal isolates were collected and examined by conventional methods for detecting S. aureus strains. PCR was used to detect S. aureus harboring the mecA and pvl genes. Out of 243 staphylococcal isolates, 55 (22.6%) S. aureus and 91 (37.4%) S. epidermidis were detected. Of the 45 patients, 21 (46.7%) were S. aureus carriers. The mecA gene was identified in 60% of S. aureus isolates, and the rest were sensitive to methicillin. Of the S. aureus isolates, 54.5% were positive for the pvl gene. This study revealed a high prevalence of PVL-positive MRSA strains in the evaluated hospitals. Thus, early diagnosis of infections caused by this pathogen seems to be necessary by intake screening allowing for proper treatment, especially in high-risk patients and in order to prevent the spread of infection.

Characterisation of clinical Staphylococcus aureus isolates harbouring mecA or Panton-Valentine leukocidin genes from four tertiary care hospitals in Indonesia.

To determine the prevalence, antimicrobial susceptibility profiles and clonal distribution of either methicillin-resistant Staphylococcus aureus (MRSA) or Panton-Valentine leukocidin (PVL)-positive S. aureus obtained from clinical cultures in Indonesian hospitals. S. aureus isolates from clinical cultures of patients in four tertiary care hospitals in Denpasar, Malang, Padang and Semarang were included. We assessed the antimicrobial susceptibility profiles using the Vitek2(®) system, determined the presence of the mecA gene and genes encoding PVL using PCR and analysed the clonal relatedness with Raman spectroscopy. SCCmec typing was performed for all MRSA isolates. Multilocus sequence typing (MLST) was performed for a subset of isolates. In total, 259 S. aureus strains were collected. Of these, 17/259 (6.6%) and 48/259 (18.5%) were MRSA and PVL-positive methicillin-susceptible S. aureus (MSSA), respectively. The prevalence of MRSA and PVL-positive MSSA ranged between 2.5-8.9% and 9.5-29.1%, respectively and depended on geographic origin. PVL-positive MRSA were not detected. Raman spectroscopy of the strains revealed multiple Raman types with two predominant clusters. We also showed possible transmission of a ST239-MRSA-SCCmec type III strain and a ST121 PVL-positive MSSA in one of the hospitals. We showed that MRSA and PVL-positive MSSA are of clinical importance in Indonesian hospitals. A national surveillance system should be set-up to further monitor this. To reduce the prevalence of MRSA in Indonesian hospitals, a bundle of intervention measures is highly recommended.

Detection of Panton-Valentine Leukocidin-Positive Methicillin-Resistant Staphylococcus aureus Nasal Carriage among Egyptian Health Care Workers.

Transmission of methicillin-resistant Staphylococcus aureus (MRSA) among patients is linked mainly to health care personnel. The Panton-Valentine leukocidin (PVL) is a cytotoxin that causes leukocyte lysis. Virulence of pvl-positive-MRSA has been attributed to its ability to express PVL toxin. Swabs for detection of nasal carriage of pvl-positive MRSA among health care personnel at Fayoum University Hospital, Fayoum, Egypt, were collected from 223 health care personnel including 70 doctors (31.4%), 95 nurses (42.6%), 21 laboratory technicians (9.4%), and 37 housekeeping staff (16.6%). Detection of MRSA was done using conventional screening methods and confirmed by multiplex polymerase chain reaction (PCR) for mecA, or its homologue mecC, and pvl genes amplification. Re-swabbing after decolonization therapy was done to evaluate the efficacy of decolonization therapy. Fifty-one of 223 participants (22.9%) were colonized with S. aureus. This included 13.5% (30/223) colonized with MRSA and 2.2% (5/223) colonized with PVL-positive MRSA. Moreover, all MRSA isolates were negative for mecC genes. Decolonization therapy was successful in 80% of MRSA carriers including all pvl-positive MRSA carriers. This is the first report on nasal carriage of pvl-positive MRSA among Egyptian health care personnel. High carriage rate of MRSA among health care personnel has been attributed mainly to poor hand hygiene compliance and non-judicious use of antibiotics. Improving compliance, reducing antibiotic overuse, screening for carriers, and decolonization are recommended strategies for reducing the spread of MRSA. Multiplex PCR could be used for confirmation of results obtained by conventional phenotypic methods.

Staphylococcus aureus Isolates Carrying Panton-Valentine Leucocidin Genes: Their Frequency, Antimicrobial Patterns, and Association With Infectious Disease in Shahrekord City, Southwest Iran

Background:A diversity of virulence factors work together to create the pathogenicity of Staphylococcus aureus. These factors include cell surface components that promote adherence to surfaces as well as exoproteins such as Panton-Valentine leukocidin (PVL), encoded by the luk-PV genes, that invade or bypass the immune system and are toxic to the host, thereby enhancing the severity of infections caused by methicillin-resistant Staphylococcus aureus (MRSA).Objectives:The aim of this study was to determine the frequency of PVL-positive MRSA strains by real-time PCR and their antibiotic susceptibility patterns by phenotypic test.Materials and Methods:In total, 284 Staphylococcus isolates, identified by phenotypic methods from clinical samples of Shahrekord University Hospitals, Shahrekord, Iran, were tested for nuc, mecA, and PVL genes by TaqMan real-time PCR. The antibiotic susceptibility patterns of PVL-containing MRSA strains were determined via the disk diffusion method.Results:In total, 196 isolates (69%) were nuc positive (i.e., S. aureus); of those isolates, 96 (49%) were mecA positive (MRSA). Eighteen (18.8%) of the 96 MRSA positive and 3 (3%) of the 100 methicillin-susceptible Staphylococcus aureus (MSSA) strains were PVL positive. PVL-positive MRSA strains were mostly recovered from tracheal specimens. Eight PVL-positive MRSA strains were resistant to all the tested antibiotics except vancomycin. A significant correlation (P = 0.001) was found between the mecA positivity and the presence of luk-PV genes.Conclusions:Community acquired (CA)-MRSA is becoming a public health concern in many parts of the world, including Asian countries. The variable prevalence of luk-PV-positive MRSA isolates in different regions and their rather high frequency in pneumonia necessitate the application of rapid diagnostic methods such as real-time PCR to improve treatment effectiveness.

Molecular epidemiology of Methicillin-resistant Staphylococcus aureus in Africa: a systematic review.

Methicillin-resistant Staphylococcus aureus (MRSA) infections are a serious global problem, with considerable impact on patients and substantial health care costs. This systematic review provides an overview on the clonal diversity of MRSA, as well as the prevalence of Panton-Valentine leukocidin (PVL)-positive MRSA in Africa. A search on the molecular characterization of MRSA in Africa was conducted by two authors using predefined terms. We screened for articles published in English and French through to October 2014 from five electronic databases. A total of 57 eligible studies were identified. Thirty-four reports from 15 countries provided adequate genotyping data. CC5 is the predominant clonal complex in the healthcare setting in Africa. The hospital-associated MRSA ST239/ST241-III [3A] was identified in nine African countries. This clone was also described with SCCmec type IV [2B] in Algeria and Nigeria, and type V [5C] in Niger. In Africa, the European ST80-IV [2B] clone was limited to Algeria, Egypt and Tunisia. The clonal types ST22-IV [2B], ST36-II [2A], and ST612-IV [2B] were only reported in South Africa. No clear distinctions were observed between MRSA responsible for hospital and community infections. The community clones ST8-IV [2B] and ST88-IV [2B] were reported both in the hospital and community settings in Angola, Cameroon, Gabon, Ghana, Madagascar, Nigeria, and São Tomé and Príncipe. The proportion of PVL-positive MRSA carriage and/or infections ranged from 0.3 to 100% in humans. A number of pandemic clones were identified in Africa. Moreover, some MRSA clones are limited to specific countries or regions. We strongly advocate for more surveillance studies on MRSA in Africa.

The incidence of Staphylococcus aureus ST8-USA300 among French pediatric inpatients is rising

Methicillin-resistant Staphylococcus aureus (MRSA) formerly colonized and infected only inpatients in hospitals, but have been reported in community settings worldwide over the last 20years. In France, the prevalence of such MRSA remains low and outbreaks have, until now, been mainly due to the ST80 clone. However, there were two outbreaks of MRSA clone ST-USA300 recently in France, including one involving children. To investigate epidemiological developments, we studied the 77 MRSA isolated from pediatric patients hospitalized between 2008 and 2013 in three French hospitals. The median incidence of MRSA was stable and low (0.137 per 100 admissions). The prevalence of Panton-Valentine leukocidin (PVL)-positive MRSA was high (33.8%). The 26 PVL-positive MRSA were genetically diverse, with two clones being predominant: ST80 (12 isolates, 46.1%) and ST8-USA300 (8 isolates, 30.8%). The incidence of ST8-USA300 increased over the 6-year period. We believe that screening for ST8-USA300 should be improved: medical biologists should be encouraged to search for PVL genes in all MRSA isolates recovered from abscesses, whatever the susceptibility pattern of the isolate, and not only when suggestive of ST80.

A 12-year survey of methicillin-resistant Staphylococcus aureus infections in Greece: ST80-IV epidemic?

Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of both healthcare-associated MRSA (HA-MRSA) and community-associated MRSA (CA-MRSA) infections. Severe MRSA infections have been associated with the virulence factor Panton–Valentine leukocidin (PVL). The aim of this study was to investigate susceptibility patterns, the presence of toxin genes, including that encoding PVL, and clonality among MRSA isolates collected from patients in Greece over a 12-year period. MRSA isolates were collected from January 2001 to December 2012 from six different hospitals. Antibiotic susceptibility was determined with the disk diffusion method and the Etest. The presence of the toxic shock syndrome toxin-1 gene (tst), the enterotoxin gene cluster (egc) and the PVL gene was tested with PCR. The genotypic characteristics of the strains were analysed by SCCmec and agr typing, and clonality was determined with pulsed-field gel electrophoresis and multilocus sequence typing. An increasing rate of MRSA among S. aureus infections was detected up to 2008. The majority of PVL-positive MRSA isolates belonged to a single clone, sequence type (ST)80-IV, which was disseminated both in the community and in hospitals, especially during the warmest months of the year. Carriage of tst was associated with ST30-IV, whereas egc was distributed in different clones. CA-MRSA isolates were recovered mainly from skin and soft tissue infections, whereas HA-MRSA isolates were associated with surgical and wound infections. During the period 2001–2012, ST80-IV predominated in the community and infiltrated the hospital settings in Greece, successfully replacing other PVL-positive clones. The predominance of ST239-III in HA-MRSA infections was constant, whereas new clones have also emerged. Polyclonality was statistically significantly higher among CA-MRSA isolates and isolates from adult patients.