USA300 methicillin-resistant Staphylococcus aureus in Zurich,Switzerland between 2001 and 2013

USA300 methicillin-resistant Staphylococcus aureus (MRSA) is the most prevalent MRSA in the United States of America (USA) and a global epidemic threat. We investigated the prevalence of USA300 at a tertiary care hospital in Zurich, Switzerland, where all MRSA strains have been collected and PFGE typed since 1992. These strains were retrospectively compared to the PFGE pattern of USA300 strain JE2. Isolates with a respective PFGE pattern were spa-typed and tested for the presence of the arginine catabolic mobile element (ACME) arc gene cluster and Panton-Valentine Leucocidin (PVL) genes. The first MRSA strain with a USA300 PFGE pattern was isolated in 2001 from a patient visiting from the USA. USA300 strains represented between 0% (in 2002) and 9.2% (in 2012) of all MRSA isolates in our hospital. We identified various USA300 subtypes based on either the PFGE pattern, the spa-type or absence of either the PVL genes or ACME arc gene cluster. All the USA300 strains including the variants (n = 47) accounted for 5.6% of all MRSA isolates typed between 2001 and 2013 and reached a maximum of 14.5% in 2009. They predominantly caused skin and soft tissue infections (74.4%). In conclusion, even though USA300 has been present in our hospital for over twelve years it has not become the predominant MRSA clone like in the USA. However, in light of the global burden of USA300, care must be taken to further contain the spread of this lineage and of MRSA in general in our hospital.     

MRSA in Austria—an overview

The aim of this study was to provide an overview of predominant and sporadic methicillin-resistant Staphylococcus aureus (MRSA) strains in large regions of Austria, and to compare the results with those from other European countries. In total, 1439 MRSA isolates, collected routinely between January 1996 and June 2006 from five Austrian federal provinces, were investigated. The isolates were confirmed as MRSA using mec A/ fem A multiplex PCR assays. Genes encoding Panton-Valentine leukocidin (PVL), which are characteristic of community-acquired MRSA, were also detected by PCR. Subtyping was performed using Sma I macrorestriction digestion of genomic DNA, followed by pulsed-field gel electrophoresis (PFGE) and cluster analysis. Isolates that could not be assigned to clusters were further analysed by spa typing and/or multilocus sequence typing. The predominant clones detected in Austria were ST228 (southern German epidemic clone), ST5 (Rhine-Hessen MRSA), the ST8 Austrian clone and CC8/ST8. Whereas the frequencies of lineages corresponding to ST247, ST45 and ST22 remained comparably low, an increase in the frequency of lineages corresponding to ST5 and to ST228 was recorded. Overall, 20 different MRSA types and 321 subtypes were recognised according to PFGE analysis. The prevalence of different strains varied considerably in the different Austrian regions. When compared to other European countries, the situation in Austria was most similar to that found in Germany.     

The molecular epidemiology and evolution of the Panton–Valentine leukocidin-positive, methicillin-resistant Staphylococcus aureus strain USA300 in Western Australia

Between 2003 and 2008, 76 clinical isolates of the Panton–Valentine leukocidin-positive Staphylococcus aureus strain 'West Australian methicillin-resistant Staphylococcus aureus (MRSA)-12' (WA MRSA-12) were recovered from 72 patients living in the Perth area in Western Australia. These isolates were found to belong to multilocus sequence type 8, and had a USA300-like pulsed-field gel electrophoresis pulsotype. All isolates were genotyped using diagnostic DNA arrays covering species markers, resistance factors, virulence-associated, as well as MSCRAMM (microbial surface components recognizing adhesive matrix molecules) genes to prove the identity between WA MRSA-12 and the pandemic strain USA300, as well as to detect possible genetic variability. In general, WA MRSA-12 isolates were similar to USA300, and the most common variant was identical to USA300- TC1516 . From this clone, most of the other variants may have evolved by a limited number of gene losses or acquisitions. Variations in carriage of virulence and resistance-associated genes allow distinction of variants or sub-clones. Altogether, 16 variants could be distinguished. They differed in the carriage of resistance genes ( blaZ/I/R , ermC , msrA + mpbBM , aadD + mupR , aphA3 + sat , tetK , qacC , merA/B/R/T ) of β-haemolysin-converting phages and of enterotoxins ( sek  +  seq , which were deleted in four isolates). Notably, the arginine catabolic mobile element (ACME) was absent in 12 isolates (15.8%). The mercury resistance ( mer ) operon, which is usually associated with SCC mec type III elements, was found in several ACME-negative isolates. The present study emphasises the importance of genotyping in detecting the introduction and evolution of significant MRSA strains within a community.     

广州市2006—2009年布鲁氏菌病发病趋势分析

目的分析广州市布鲁氏菌病（简称布病）疫情,了解布病疫情的流行特点和发展趋势,为今后制定防治措施提供依据。方法利用“国家疾病监测信息报告管理系统”报告的病例数和监测中的数据,进行流行病学分析。结果2006—2009年,广州市共报告布病31例,其中男20例,女11例,年龄最大69岁,最小19岁,以青壮年居多;病例几乎都与活羊、羊肉和羊内脏接触有关;临床表现以发热、多汗、关节游走性疼痛为主。结论来自布病疫区感染布病的羊只流人我市、从业人员缺乏防病意识、检疫和免疫措施不到位是造成我市布病发病数上升的主要原因。因此,加强人市牲畜检疫和从业人员布病健康教育是当前重要的策略。     

PCR for the identification of methicillin-resistant Staphylococcus aureus (MRSA) strains using a single primer pair specific for SCCmec elements and the neighbouring chromosome-borne orfX

The chromosomal location of the SCCmec elements containing mecA allows the identification of methicillin-resistant Staphylococcus aureus (MRSA) strains by PCR amplification of a sequence covering the right junction of the SCCmec elements and the adjacent chromosomal region encoding the species-specific ORFX. MRSA strains can be identified specifically using one forward primer, with only one or two mismatches, targeting the SCCmec elements of different types, and one reverse primer targeting the orfX region.     

The High Diversity of MRSA Clones Detected in a University Hospital in Istanbul

Background: To characterize the methicillin-resistant Staphylococcus aureus (MRSA) clones present in Istanbul, 102 MRSA isolates collected during a 5-year period at the Istanbul Medical Faculty Hospital were characterized using microarray analysis and phenotypic resistance profiles.Methods: Resistance to methicillin was detected with a cefoxitin disk diffusion assay and confirmed with a MRSA-agar and MRSA detection kit. Antimicrobial susceptibility testing was performed by a disk diffusion assay and interpreted according to the 2012 guidelines of the Antibiogram Committee of the French Society for Microbiology. Decreased susceptibility to glycopeptides was confirmed using the population analysis profile-area under the curve (PAP-AUC) method. The presence of the mecA gene was detected by polymerase chain reaction. Bacterial DNA was extracted according to the manufacturer''s recommended protocol using commercial extraction kits. Strains were extensively characterized using the DNA microarray.Results: Isolates were grouped into six clonal complexes. The most frequently detected clone was the Vienna/Hungarian/Brazilian clone (ST239-MRSA-III), which accounted for 53.9% of the isolates. These isolates were resistant to multiple antibiotics, particularly penicillin, tetracycline, rifampicin, kanamycin, tobramycin, gentamicin, levofloxacin, erythromycin, lincomycin and fosfomycin. Furthermore, three isolates were detected by population analysis profile as heterogeneous vancomycin-intermediate S. aureus (hVISA). The UK-EMRSA-15 clone (ST22-MRSA-IV PVL negative) was detected in 9.8% of the isolates and was mainly susceptible to all anti-staphylococcal antibiotics. Seven isolates (6.9%) were positive for PVL genes and were assigned to the CC80-MRSA-IV clone (European CA-MRSA clone, three isolates), ST8-MRSA-IV clone (USA300 clone, two isolates, one ACME-positive) or ST22-MRSA-IV clone (“Regensburg EMRSA” clone, two isolates). All other clones were detected in one to six isolates and corresponded to well-known clones (e.g., Pediatric clone, Dublin EMRSA clone, WA MRSA-54/63, WA MRSA-1/57).Conclusions: This work highlighted both the high prevalence of ST239-MRSA-III clone and the large diversity of the other MRSA clones detected in a university hospital in Istanbul.     

A variant of the Southern German clone of methicillin-resistant Staphylococcus aureus is predominant in Croatia

The aim of the present study was to investigate the antibiotic susceptibility patterns and molecular epidemiology of clinical methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered in 24 hospitals in 20 cities in Croatia from October to December 2004. A total of 1815 consecutive S. aureus isolates were recovered, 248 of which were MRSA. The MRSA isolates were analysed using spa typing, multilocus sequence typing and SCCmec typing. Furthermore, the presence of Panton–Valentine leukocidin (PVL) genes was determined as a genetic marker for community-associated MRSA. The MRSA prevalence was 14%. Ninety-six per cent of the MRSA isolates were resistant to ciprofloxacin, 95% to clindamycin and azithromycin, 94% to gentamicin, and 93% to erythromycin. The majority of the MRSA isolates (78%) was associated with the ST111-MRSA-I clone. In addition, various other endemic MRSA clones were observed, such as the ST247-MRSA-I (4%), the ST45-MRSA-IV (2%), the ST5-MRSA-I (2%), the ST239-MRSA-III (2%), the ST5-MRSA-II (1%), the ST8-MRSA-IV (1%) and the ST5-MRSA-IV (<1%) clones. Furthermore, we observed one PVL-negative ST80-MRSA-IV isolate. Four PVL-positive MRSA isolates were found, associated with ST8-MRSA-IV, ST80-MRSA-IV and ST80-MRSA-I. The ST111-MRSA-I clone was predominant in Croatia. Future surveillance studies of MRSA are important to elucidate whether changes in the clonal distribution of MRSA will occur, and if the minor endemic MRSA clones observed in the present study will replace the ST111-MRSA-I clone on a large scale.     

Significance of MRSA strains in community associated skin and soft tissue infections

The study was conducted to determine the antibiotic susceptibility profile of community-associated methicillin resistant Staphylococcus aureus (CAMRSA) strains isolated from infections. S. aureus strains were isolated from clinical specimens using the standard procedures. CDC definition was used to classify CAMRSA. Antibiotic susceptibility test was done using Kirby-Bauer disk diffusion method. Double disk diffusion method (D-test) was used to detect inducible macrolide, lincosamide and streptogramin_B resistance (inducible MLS_B resistance). A total of 83 CAMRSA were isolated from abscesses and other skin infections in persons without known risk factors for MRSA infection. All CAMRSA were susceptible to vancomycin. Out of 83 CAMRSA, 13 (15.65%) were D-test positive (inducible MLS_B positive) and 6 (7.23%) were positive for constitutive MLS_B resistance. Eight strains (9.63%) were resistant to tetracycline and 26 (31.32%) strains were resistant to erythromycin. Increased rate of inducible clindamycin resistance among CAMRSA indicates the importance of identification of such strains by D test to avoid treatment failure when clindamycin is used.     

Molecular epidemiology of predominant clones and sporadic strains of methicillin resistant Staphylococcus aureus in Switzerland and comparison with European epidemic clones

Objective   To assess the molecular epidemiology and risk factors of predominant clones and sporadic strains of methicillin-resistant Staphylococcus aureus (MRSA) in Swiss hospitals and to compare them with European strains of epidemic clones.
Material and methods   One-year national survey of MRSA cases. Analysis of epidemiological and molecular typing data (PFGE) of MRSA strains.
Results   In 1997, 385 cases of MRSA were recorded in the five Swiss university hospitals and in 47 community hospitals. Half of the cases were found in Geneva hospitals where MRSA was already known to be endemic. Molecular typing of 288 isolates (one per case) showed that 186 (65%) belong to four predominant clones, three of which were mostly present in Geneva hospitals. In contrast, the fourth clone (85 cases) was found in 23 hospitals (in one to 16 cases per hospital). The remaining 35% of the strains were clustered into 62 pulsed field gel electrophoresis types. They accounted for one to five patients per hospital and were defined as sporadic. Multivariate analysis revealed no independent risk factors for harboring a predominant versus a sporadic strain, except that transfer from a foreign hospital increases the risk of harboring a sporadic strain (OR, 42; 95% CI, 5–360).
Conclusion   While cases with predominant clones were due to the local spread of these clones, most sporadic cases appear to be due to the continuous introduction of new strains into the country. With the exception of a transfer from a hospital outside Switzerland, no difference in the clinical or epidemiological characteristics was observed between patients harboring a predominant clone and those with a sporadic strain.     

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.     

汕头大学医学院附属第一医院耐甲氧西林金黄色葡萄球菌(MRSA)耐药谱分析

目的:了解汕头大学医学院附属第一医院分离的92株耐甲氧西林金黄色葡萄球菌(MRSA)的耐药情况。方法:琼脂稀释法测定17种抗菌药物对MRSA的最低抑菌浓度(MIC)。结果:92株MRSA对实验中所有β-内酰胺类抗生素的耐药率均在80%以上。绝大部分菌株对庆大霉素、红霉素、氟喹诺酮类也不敏感。超过半数的MRSA菌株对利福平保持敏感,对氯霉素、多西环素及米诺环素也普遍敏感,未发现耐万古霉素的MRSA菌株。结论:我院附属第一医院的MRSA呈多重耐药,但对氯霉素,半合成四环素及万古霉素仍然敏感。     

Failure of bacteriophage typing to detect an inter-hospital outbreak of methicillin-resistant Staphylococcus aureus (MRSA) in Zagreb subsequently identified by random amplification of polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE)

Objective: To establish the extent of inter-hospital spread of methicillin-resistant Staphylococcus aureus (MRSA) in Zagreb and to determine the most suitable method for typing local strains.
Methods: We analyzed a collection of 33 MRSA isolates from three Zagreb hospitals together with five unrelated British MRSA isolates by antibiogram typing, bacteriophage typing, randomly amplified polymorphic DNA (RAPD) analysis and pulsed-field gel electrophoresis (PFGE) after digestion with Smal restriction endonuclease. Bacteriophage typing was done with the international set of S. aureus typing phages. RAPD and PFGE profiles were analyzed visually and by using the 'GelCompar' computer program.
Results: Antibiogram typing provided eight profiles. Thirty (91%) of the 33 Croatian strains of MRSA were non-typable by phage typing. Visual analysis of RAPD products identified six, and visual analysis of PFGE fragments nine, distinct profiles. Computer analysis of RAPD data separated British isolates from the Croatian ones, but did not cluster the visually determined RAPD types. PFGE computer analysis separated British isolates and clustered isolates in concordance with visual interpretation. Thirty-one of the 38 isolates (82%) were visually grouped in the same clusters by both molecular methods. The dominant strain was present in each of the three hospitals.
Conclusions: Bacteriophage typing was unhelpful for the analysis of Croatian MRSA, since most strains were untypable with the international set of bacteriophages. RAPD and PFGE were more successful in typing the organisms and showed evidence of inter-hospital spread of one predominant MRSA strain in all three Zagreb hospitals. Thus RAPD and PFGE proved to be a useful aid in elucidating the epidemiology of MRSA infection in Zagreb hospitals and should be established in Croatia for typing MRSA.     

Treatment of MRSA infections in India: Clinical insights from a Delphi analysis

PurposeInternational and Indian guidelines for the management of patients with methicillin-resistant Staphylococcus aureus (MRSA) infections are available, but the local guidelines are not MRSA-specific. This study aimed to provide clinical insights for the treatment of MRSA infections in India.MethodsWe used a three-step modified Delphi method to obtain insights. Ten experts comprising infectious disease specialists, microbiologists, pulmonologists, and critical care experts agreed to participate in the analysis. In round 1, a total of 161 statements were circulated to the panel and the experts were asked to ‘agree’ or ‘disagree’ by responding ‘yes’ or ‘no’ to each statement and provide comments. The same process was used for 73 statements in round 2. Direct interaction with the experts was carried out in round 3 wherein 35 statements were discussed. At least 80% of the experts had to agree for a statement to reach concordance.ResultsEighty-eight statements in round 1, thirty-eight statements in round 2, and eight statements in round 3 reached concordance and were accepted without modification. The final document comprised 152 statements on the management of various syndromes associated with MRSA such as skin and soft tissue infections, bacteremia and endocarditis, pneumonia, bone and joint infections, and central nervous system infections.ConclusionsThis analysis will assist clinicians in India to choose an appropriate course of action for MRSA infections.     

The use of spa and phage typing for characterization of a MRSA population in a Belgian hospital: Comparison between 2002 and 2007

Target of the study

Strain typing of pathogens is essential to pinpoint the sources and routes of transmission and to forecast future trends. In a general hospital, we studied possible changes in the MRSA population.

Patients and methods

MRSA isolates received from a Belgian general hospital, during 2002 (n = 150) and the second half of 2007 (n = 105), were compared by phage and spa typing.

Results

In 2002, [J]* phage types characterized 45% of the MRSA isolates, 13% belonged to the [O]* phage types, 12% to a local phage type 29/42E/54/D11* and 28% were not assigned to a defined group. Thirteen different spa types were found among the isolates: 39% belonged to t038, 27% to t121, 14% to t041, 5% to t740, and 4% to t002 and t024 each. Two spa types were found respectively in two and three isolates, five were unique.In 2007, 35% belonged to [J]*, 23% to [O]* and 39% could not be put in a defined group. Eighteen different spa types were found: 30% belonged to t740, 29% to t121, 13% to t038 and 10% to t002. Three spa types were represented in two isolates, eleven were unique.The t041 spa type was specific for the 29/42E/54/D11* and the majority of the t121 isolates were related to [J]*.

Conclusion

[J]* remained the dominant phage types group but decreased whereas [O]*, the second phage types group, increased. As to the spa types, t740 became dominant while t121 remained second. Phage and spa typing point to some quantitative changes among the Belgian MRSA population.     

Emergence and control of an outbreak of infections due to Panton-Valentine leukocidin positive,ST22 methicillin-resistant Staphylococcus aureus in a neonatal intensive care unit

Methicillin resistant Staphylococcus aureus (MRSA) infection can cause significant morbidity and mortality in neonates. We investigated a nosocomial MRSA outbreak in a neonatal intensive care unit (NICU), using a novel typing method. Following two fatal cases, in May 2011, a prospective outbreak investigation was conducted, involving neonates, mothers and healthcare workers in a large tertiary NICU in Sydney. MRSA isolates were characterized by antimicrobial susceptibility testing, a multiplex PCR-based reverse line blot (mPCR/RLB) binary typing system and other molecular typing methods. Over 7 months, 14 neonates were colonized with MRSA and six infected: three with superficial lesions and three with life-threatening disease, including the two index cases, who died despite empirical treatment with vancomycin. Isolates from 15 neonates were indistinguishable by RLB typing and identified as a PVL-producing ST22 SCCmec IV MRSA strain, which was resistant to gentamicin and trimethoprim-sulphamethoxazole. The outbreak strain was also isolated from one healthcare worker, one environmental swab and one father, but the source remained obscure. During the same period several different non-multiresistant and multiresistant MRSA strains were isolated from five neonates, five mothers (including two whose infants were colonized with the outbreak strain), one father, three healthcare workers and two environmental swabs. Rapid turnaround time of typing results allowed us to recognize and define the outbreak and implement targeted infection control interventions. PVL-producing ST22 SCCmec IV MRSA appears to be a virulent and highly transmissible pathogen in the NICU, which was difficult to control.     

Molecular and phenotypic characterization of invasive group B streptococcus strains from infants in Norway 2006–2007

Multilocus sequence typing of an almost complete collection of invasive group B streptococcus (GBS) strains from infants in Norway, conducted in 2006–2007, revealed 27 sequence types (ST), of which 23 clustered into five clonal complexes. The case fatality rate of invasive GBS disease in infants was 16/98 (16.3%). Type V strains were predominant among strains resistant to erythromycin and clindamycin (11/18; 61.1%). All type V strains from fatal cases (5/16) were ST1, resistant to erythromycin and clindamycin, and belonged to three pulsed-field gel electrophoresis-clusters. Further analysis of virulence characteristics of these apparently highly virulent subtypes of type V, ST1 GBS strains is warranted.     

Clinical isolates of Staphylococcus aureus from the Arkhangelsk region,Russia: antimicrobial susceptibility,molecular epidemiology,and distribution of Panton‐Valentine leucocidin genes

A total of 91 consecutive clinical isolates of Staphylococcus aureus were collected at the Regional Hospital of Arkhangelsk, Russia, from May to December 2004, and examined for antimicrobial susceptibility, methicillin resistance and presence of Panton‐Valentine leucocidin (PVL) genes. Epidemiological typing was performed by pulsed‐field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Methicillin‐resistant S. aureus (MRSA) isolates were examined by staphylococcal cassette chromosome mec (SCCmec) typing. High‐to‐moderate rates of resistance to penicillin (β‐lactamase production; 93%), tetracycline (40%), erythromycin and clindamycin (32%) were observed. Forty out of ninety‐one (44%) isolates were positive for PVL genes. Thirty‐six (40%) PVL‐positive methicillin‐susceptible S. aureus (MSSA) strains were shown by PFGE and MLST typing (ST121, ST681, ST837) to be part of a nosocomial outbreak caused by clonal complex (CC) 121. PFGE, MLST and SCCmec typing revealed three MRSA clones. Sequence type (ST) 239‐III (n=11), ST1097‐III (n=1) and ST8‐IV (n=3) belong to CC8 of epidemic multiresistant MRSA, whereas ST426‐MRSA‐IV/CC395 (n=1) has not been reported previously. All MRSA strains were PVL negative. The overall results underline the necessity of microbiological sampling, antimicrobial susceptibility testing, and epidemiological typing as a rational basis for antimicrobial treatment of S. aureus infections, and infection control measures to limit the spread of multiresistant MRSA and epidemic MSSA clones.