Superantigen Gene Profiles Research Articles

Interactions between staphylococcal enterotoxins A and D and superantigen-like proteins 1 and 5 for predicting methicillin and multidrug resistance profiles among Staphylococcus aureus ocular isolates.

BackgroundMethicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant (MDR) S. aureus strains are well recognized as posing substantial problems in treating ocular infections. S. aureus has a vast array of virulence factors, including superantigens and enterotoxins. Their interactions and ability to signal antibiotics resistance have not been explored.ObjectivesTo predict the relationship between superantigens and methicillin and multidrug resistance among S. aureus ocular isolates.MethodsWe used a DNA microarray to characterize the enterotoxin and superantigen gene profiles of 98 S. aureus isolates collected from common ocular sources. The outcomes contained phenotypic and genotypic expressions of MRSA. We also included the MDR status as an outcome, categorized as resistance to three or more drugs, including oxacillin, penicillin, erythromycin, clindamycin, moxifloxacin, tetracycline, trimethoprim-sulfamethoxazole and gentamicin. We identified gene profiles that predicted each outcome through a classification analysis utilizing Random Forest machine learning techniques.FindingsOur machine learning models predicted the outcomes accurately utilizing 67 enterotoxin and superantigen genes. Strong correlates predicting the genotypic expression of MRSA were enterotoxins A, D, J and R and superantigen-like proteins 1, 3, 7 and 10. Among these virulence factors, enterotoxin D and superantigen-like proteins 1, 5 and 10 were also significantly informative for predicting both MDR and MRSA in terms of phenotypic expression. Strong interactions were identified including enterotoxins A (entA) interacting with superantigen-like protein 1 (set6-var1_11), and enterotoxin D (entD) interacting with superantigen-like protein 5 (ssl05/set3_probe 1): MRSA and MDR S. aureus are associated with the presence of both entA and set6-var1_11, or both entD and ssl05/set3_probe 1, while the absence of these genes in pairs indicates non-multidrug-resistant and methicillin-susceptible S. aureus.ConclusionsMRSA and MDR S. aureus show a different spectrum of ocular pathology than their non-resistant counterparts. When assessing the role of enterotoxins in predicting antibiotics resistance, it is critical to consider both main effects and interactions.

018 TSST-1+ Staphylococcus aureus in bullous pemphigoid

Bullous pemphigoid (BP) is an autoimmune blistering disease that is treated with high dose immunosuppression due to lack of specific targets. Staphylococcus aureus is a commensal bacterium implicated inflammatory and autoimmune disorders because of its secretion of toxins with superantigen effects. We prospectively evaluated S. aureus colonization and its production of toxic shock syndrome toxin-1 (TSST-1) in 28 new onset BP patients. Inclusion criteria were active blistering and linear basement membrane IgG/C3 or a serum ELISA >14 for BP180 IgG. Bacterial swabs were obtained from the lesion interior, nares and unaffected skin of BP patients and nares and anatomically matched skin of 28 age- and sex-matched controls. Staphylococcal growth was assessed on blood agar, and TSST-1 production by cultured S. aureus isolates and in blister fluid was evaluated by immunoblot. S. aureus colonization of BP lesions was 3-6-fold higher than the nares or unaffected skin from the same patients (p≤0.300) and 6-fold higher than control nares or skin (p≤0.0015). Evaluation of superantigen gene profiles using PCR indicated that 96% of BP patients are colonized with a clonal strain of TSST-1+ S. aureus. Colonization and circulating levels of TSST-1 neutralizing antibodies, measured by ELISA, did not correlate. Interestingly, S. aureus colonization was not observed in patients who had received prior antibiotics. In colonized patients with severe disease, addition of anti-staphylococcal antibiotics resulted in clinical improvement and eliminated lesional colonization. This study shows that BP lesions harbor a clonal strain of TSST-1 producing S. aureus that is not evident in the general elderly population. Thus, immunosuppressive therapies should be balanced with the knowledge that S. aureus is likely present in BP lesions and the knowledge that antibiotics may play an important therapeutic role through bacterial clearance.

Detection of Group A Beta Hemolytic Streptococci Species, emm, and Exotoxin Genes Isolated from Patients with Tonsillopharyngitis.

Group A Beta Hemolytic Streptococci (GAS) is the most critical human pathogen that leads to tonsillopharyngitis. The aims of this study were to identify GAS isolates and to determine emm typing, the coverage rate of available vaccines, and the distribution of superantigen gene profiles. 15 GAS isolates were isolated from throat cultures of 200 patients with tonsillopharyngitis, who were admitted to Canakkale Health Application and Research Hospital between October 2017 and May 2018. Identification of the isolates was performed by conventional methods and 16S rRNA sequence analysis. emm typing and exotoxin profiling of the isolates were performed by polymerase chain reaction. 7.5% GAS was detected in 200 patients. All the GAS isolates were identified as S. pyogenes. emm typing can be carried out in 13 S. pyogenes isolates. emm89 (33.3%), emm44 (20%), emm6 (13.3%), emm84 (6.7%), emm1 (6.7%), and emm18.1 (6.7%) were found to be six emm types. The coverage rate of S. pyogenes strains for 26-valent vaccine was 61.5% and for the 30-valent vaccine 84.6%. The most common exotoxin was speB (86.7%), followed by speC (60%), speF (33.3%), ssa (26.7%), speA (20%), speM (20%), speJ (13.3%), speL (6.7%), and speI (6.7%). As a result of determining the emm types of S. pyogenes species in Canakkale, it was concluded that the potential of 30-valent vaccine should be considered in Turkey and development of vaccines containing exotoxin types may be beneficial.

Molecular characteristics and antimicrobial susceptibility of Staphylococcus aureus among children with respiratory tract infections in southwest China.

The molecular characteristics and antimicrobial susceptibility of Staphylococcus aureus (S. aureus) in general pediatric wards and county-level hospitals were rarely reported in China. Staphylococcus aureus was isolated from children hospitalized with respiratory tract infection (RTI) in Zhongjiang and Youyang counties in 2015. All isolates were typed by multilocus sequence, staphylococcal protein A, accessory gene regulator (agr), and staphylococcal cassette chromosome mec [SCCmec, for methicillin-resistant S. aureus (MRSA) only]. Polymerase chain reaction was used to screen 21 super-antigen (SAg) genes and panton-valentine leukocidin (pvl). Antimicrobial susceptibility testing was performed by E test. A total of 2136 children were enrolled. Overall, 125 (5.9%) children carried S. aureus, among which MRSA accounted for 42.4%. ST59-SCCmec type IV-t437-agr group I (58.5%) was the most prevalent genotype in MRSA, and ST188-t189-agr group I (22.2%) was the top genotype in methicillin-sensitive S. aureus (MSSA). The pvl carriage rate in MRSA and MSSA was 15.1% and 9.7%, respectively (P = 0.4112). About 96.8% of S. aureus isolates were positive for at least one SAg gene. The most common SAg gene profile in the dominant ST59 clone was seb-sek-seq (42.8%). All S. aureus isolates were resistant to penicillin and erythromycin (minimum inhibitory concentration 90 was > 32 and 256mg/L to penicillin and erythromycin, respectively), but usually susceptible to other tested non-β-lactam antimicrobials. Staphylococcus aureus and MRSA were detected with a high frequency in children with RTI in county-level hospitals of China. ST59-SCCmec type IV-t437-agr group I was the dominant MRSA clone. The S. aureus isolates exhibited high resistance to penicillin and erythromycin.

Potential role of host defense antimicrobial peptide resistance in increased virulence of health care-associated MRSA strains of sequence type (ST) 5 versus livestock-associated and community-associated MRSA strains of ST72.

The most significant community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) in Korea is sequence type (ST) 72 with staphylococcal cassette chromosome mec (SCCmec) type IV (ST72-MRSA-IV). Although the impact of CA-MRSA on the clinical outcomes versus healthcare-associated (HA)-MRSA remains unclear, it has recently been revealed that ST5 HA-MRSA-II is associated with higher mortality compared with ST72 CA-MRSA-IV, suggesting higher virulence in ST5 HA-MRSA-II strains. In this investigation, human-/animal-originated ST72-MRSA-IV strains were examined for virulence phenotypes and compared with those of ST5-MRSA-II strains, the established HA-MRSA in Korea. Overall, ST5 HA-MRSA-II strains demonstrated higher levels of resistance to host defense-cationic antimicrobial peptides of human (LL-37), bovine (BMAP-28), and bacterial (polymyxin B) origins versus ST72-MRSA-IV strains via enhanced surface positive charge. Hemolysis profiles, gelatinase activity, and staphylococcal superantigen gene profiles were not different between ST72 CA-MRSA and ST5 HA-MRSA strains. However, ST5 HA-MRSA strains were able to downregulate initial cytokine response in murine macrophages.

Streptococcus pyogenes Causing Skin and Soft Tissue Infections Are Enriched in the Recently Emerged emm89 Clade 3 and Are Not Associated With Abrogation of CovRS.

Although skin and soft tissue infections (SSTI) are the most common focal infections associated with invasive disease caused by Streptococcus pyogenes (Lancefield Group A streptococci - GAS), there is scarce information on the characteristics of isolates recovered from SSTI in temperate-climate regions. In this study, 320 GAS isolated from SSTI in Portugal were characterized by multiple typing methods and tested for antimicrobial susceptibility and SpeB activity. The covRS and ropB genes of isolates with no detectable SpeB activity were sequenced. The antimicrobial susceptibility profile was similar to that of previously characterized isolates from invasive infections (iGAS), presenting a decreasing trend in macrolide resistance. However, the clonal composition of SSTI between 2005 and 2009 was significantly different from that of contemporary iGAS. Overall, iGAS were associated with emm1 and emm3, while SSTI were associated with emm89, the dominant emm type among SSTI (19%). Within emm89, SSTI were only significantly associated with isolates lacking the hasABC locus, suggesting that the recently emerged emm89 clade 3 may have an increased potential to cause SSTI. Reflecting these associations between emm type and disease presentation, there were also differences in the distribution of emm clusters, sequence types, and superantigen gene profiles between SSTI and iGAS. According to the predicted ability of each emm cluster to interact with host proteins, iGAS were associated with the ability to bind fibrinogen and albumin, whereas SSTI isolates were associated with the ability to bind C4BP, IgA, and IgG. SpeB activity was absent in 79 isolates (25%), in line with the proportion previously observed among iGAS. Null covS and ropB alleles (predicted to eliminate protein function) were detected in 10 (3%) and 12 (4%) isolates, corresponding to an underrepresentation of mutations impairing CovRS function in SSTI relative to iGAS. Overall, these results indicate that the isolates responsible for SSTI are genetically distinct from those recovered from normally sterile sites, supporting a role for mutations impairing CovRS activity specifically in invasive infection and suggesting that this role relies on a differential regulation of other virulence factors besides SpeB.

Prevalent emm types and superantigen gene patterns of group A Streptococcus in Thailand.

Group A Streptococcus (GAS) are globally distributed bacterial pathogens. We examined the emm genotypes, which are important indicators of virulence, of 349 clinical GAS isolates collected using two surveillance systems, i.e. Invasive Bacterial Infection Surveillance (IBIS) from 2010 to 2011 (234 isolates) and routine surveillance of clinically isolated bacteria from various hospitals during 1996-2011 (115 isolates) in Thailand. The major emm genotypes in IBIS samples were emm44 (12·0%), emm104 (6·8%), emm22 (5·6%), and emm81 (5·6%), whereas only one isolate (0·4%) had the emm1 genotype, which is significantly more common in invasive cases in the Western world. In samples collected during routine surveillance, emm238 (10·4%), emm44 (8·7%), and emm165 (7·0%) were dominant. The major superantigen gene profiles were similar between the groups, and 30·1% of isolates did not possess the phage-encoded superantigens (speA, speC, speH, speI, speK, speL, speM, ssa). Although most isolates exhibited limited gene profiles, emm44 isolates had highly variable gene profiles (15 patterns). We conclude that emm44 is the predominant GAS genotype in Thailand, and isolates varied in superantigen gene profiles.

Superantigen Gene Profile of Staphylococcus Aureus Colonizing Egyptian Pediatric Patients with Atopic Dermatitis : Relationship to Disease Severity and Multi-Drug Resistance

Background: Eczematous skin of atopic dermatitis (AD) is highly susceptible to infection and colonization by Staphylococcus aureus and the superantigen toxins can worsen the condition. Objectives: To assess the colonization of Egyptian pediatric AD patients with S. aureus and to characterize the superantigen gene profile of isolates in relation to severity and to presence of multiple drug resistant (MDR) strains. Methodology: The study included 53 AD pediatric patients and 45 controls. Severity of AD was assessed by scoring atopic dermatitis (SCORAD) index. Swabs were collected to isolate S. aureus. Isolates were subjected to multiplex PCR reactions for detection of six superantigen genes and to antimicrobial susceptibility tests by disc diffusion method. Results: Colonization with S. aureus was significantly higher (P < 0.0001) in AD children compared to controls and was significantly associated (P= 0.001) with severity. Superantigen genes were detected in 30.1% of isolates. The most prevalent genes were sea (64.5%), seb (32.3%), sec (6.5%) and tsst-1 (3.2%). Multidrug resistance was found in 63.1% of strains. Severity of AD was significantly higher with strains harboring superantigen genes (P=0.04) and with MDR strains (P=0.0002). Among methicillin resistant S. aureus (MRSA), seb was the most prevalent superantigen gene (37.5%), while sea was most prevalent in methicillin-susceptible S. aureus (MSSA) (20%), MDR (23.1%) and non MDR isolates (13.2%).Conclusion: Superantigen genes and multidrug resistance are common in S. aureus colonizing AD patients and are associated with severity. More attention should be paid at performing antimicrobial susceptibility testing before antibiotic therapy.

Superantigen gene profiles, genetic relatedness and biological activity of exosecretions of Staphylococcus aureus isolates obtained from milk of cows with clinical mastitis

This study evaluated the superantigen gene profiles, genetic relatedness and biological activity of exosecretions of 50 Staphylococcus aureus isolates obtained from milk of cows with clinical mastitis. Genomic relatedness of S. aureus was determined by pulsed field gel electrophoresis analysis of macro-restricted chromosomes. The presence of genes encoding superantigens was confirmed by multiplex PCR. To study the biological activity of S. aureus exosecretions, the supernatants from bacterial liquid cultures were classified into three groups: those with leukotoxin‐like properties, those with superantigen‐like properties and those with no particular activity on leukocytes cultured in vitro. It was shown that all analyzed bacterial isolates belonged to the same clonal type and harbored the same combination of superantigen genes, namely sed, selj and ser. However, 22% of all isolates produced factors with superantigen‐like and 48% of them with leukotoxin‐like activities. Finally, although there were no detectable genetic differences between the analyzed bacterial isolates, the virulence factors secreted by them differed considerably.

Comparative analysis of emm types, superantigen gene profiles and antibiotic resistance genes among Streptococcus pyogenes isolates from ocular infections, pharyngitis and asymptomatic children in south India

Group A Streptococcus (Streptococcus pyogenes) is responsible for a wide array of infections and incidence is high in developing countries like India. Although distribution of emm types of S. pyogenes in India has been described, its association with the virulence genes and ocular isolates is less concentrated. In the present study emm type surveillance as well as its association with toxin gene profile was analyzed. Ocular infected cases such as lacrimal abscess, corneal ulcers, mucocoele showed the presence of 20 S. pyogenes isolates. For noninvasive isolates, we screened 370 pharyngitis cases and 400 asymptomatic school children and recovered 33 pharyngitis and 14 carrier isolates respectively. 14 Emm type distributions were observed in ocular isolates, 11 emm types each in pharyngitis and asymptomatic carrier isolates. The two dominant emm types, emm49 and emm63 were accounted for 33% of the total S. pyogenes isolates. Among ocular isolates, slo, smeZ, speB and speG were found in >50% of isolates, in pharyngitis smeZ (48%), speB (45%) and speG (42%) genes were found to be prevalent. Alarmingly, carrier isolates showed more prevalence to virulence genes than the ocular and pharyngitis isolates with speF (79%), speB, speG (64%), slo and sil (64%). Among the three groups, pharyngitis isolates harbored more prtF1 (33%) and prtF2 (94%) than the asymptomatic carriers (28% and 71%) and the ocular isolates (45% and 40%). 450bp Size band in prtF1 and 350bp size band in prtF2 showed dominance. Among the three groups tested, the distribution of ermB and mefA was high in pharyngitis isolates (30%) where 10 isolates showed the presence of both genes. None of the isolates showed the presence of ermA and tetO genes. Dendrogram generated based on the virulence and antibiotic resistance gene profiles revealed that except one cluster, all other clusters showed some correlation with ocular, pharyngitis and asymptomatic carrier isolates, irrespective of their emm types.

Superantigen gene complement of Streptococcus pyogenes—relationship with other typing methods and short-term stability

The profiling of the superantigen (SAg) encoding genes has been frequently used as a complementary typing method for group A streptococci (GAS), but a confusing gene nomenclature and a large diversity of primers used in screening has led to some conflicting results. The aim of this work was to develop a polymerase chain reaction (PCR) method capable of efficiently amplifying all the known allelic variants of these genes, and to evaluate the congruence of this methodology with other commonly used molecular typing methods. The presence of the 11 known SAg genes and two other exotoxin-encoding genes (speB and speF) was tested in a collection of 480 clinical GAS isolates, using two multiplex PCR reactions. The SAg gene profile was compared with other typing methods. Four naturally occurring deletions involving the genes speB, speF, and rgg were characterized, two of which were found among invasive isolates. The absence of the chromosomally encoded genes speG and smeZ was supported by Southern blot hybridization and associated with specific GAS lineages, while the presence of phage-encoded genes was more variable. Positive associations between SAg genes or between SAg profiles and emm types or pulsed-field gel electrophoresis (PFGE) clusters were observed. The results suggest that the SAg profile diversifies faster than other properties commonly used for molecular typing, such as emm type and multilocus sequence typing (MLST) sequence types (STs), and can be a useful complement in GAS molecular epidemiology. Still, the short-term stability of the SAg gene profile among prevalent genetic lineages may largely explain the observed associations between SAg genes.

Unique distribution of emm types and superantigen gene profiles of group A streptococci isolated in Thailand

Unique distribution of emm types and superantigen gene profiles of group A streptococci isolated in Thailand

Clinical and Microbiologic Characteristics of Invasive Streptococcus pyogenes Infections in North and South India

The lack of epidemiologic data on invasive Streptococcus pyogenes infections in many developing countries is concerning, as S. pyogenes infections are commonly endemic in these areas. Here we present the results of the first prospective surveillance study of invasive Streptococcus pyogenes infections in India. Fifty-four patients with invasive S. pyogenes infections were prospectively enrolled at two study sites, one in the north and one in the south of India. Sterile-site isolates were collected, and clinical information was documented using a standardized questionnaire. Available acute-phase sera were tested for their ability to inhibit superantigens produced by the patient's own isolate using a cell-based neutralizing assay. The most common clinical presentations were bacteremia without focus (30%), pneumonia (28%), and cellulitis (17%). Only two cases of streptococcal toxic shock syndrome and no cases of necrotizing fasciitis were identified. Characterization of the isolates revealed great heterogeneity, with 32 different emm subtypes and 29 different superantigen gene profiles being represented among the 49 sterile-site isolates. Analyses of acute-phase sera showed that only 20% of the cases in the north cohort had superantigen-neutralizing activity in their sera, whereas 50% of the cases from the south site had neutralizing activity. The results demonstrate that there are important differences in both clinical presentation and strain characteristics between invasive S. pyogenes infections in India and invasive S. pyogenes infections in Western countries. The findings underscore the importance of epidemiologic studies on streptococcal infections in India and have direct implications for current vaccine developments.

Similar Superantigen Gene Profiles and Superantigen Activity in Norwegian Isolates of Invasive and Non‐Invasive Group A Streptococci

Group A streptococcus (GAS) harbours several virulence factors, including M protein (coded by the emm gene) and superantigens (SAgs). SAgs are extracellular toxins that directly activate the immune system by cross-binding to the HLA class II molecule and T cell receptor (TCR), thereby causing activation of up to 30% of the T cells and subsequent massive secretion of cytokines. Forty-eight GAS strains isolated from patients at Norwegian hospitals between 1988 and 2004 were included in this study. Of these, 24 were invasive streptococcal toxic shock syndrome (STSS) or necrotizing fasciitis (NF) isolates and 24 were non-invasive pharyngitis isolates, matched for having the same T-type and year of isolation as the invasive isolates. The isolates were characterized by emm sequence typing, multilocus sequence typing (MLST) and SAg gene profiles. A correlation between T-type, emm type, sequence type and SAg gene profile was revealed. No difference between invasive and non-invasive isolates regarding serotype or genotype was demonstrated. Selected invasive and non-invasive isolates with identical SAg gene profiles were analysed for SAg activity in bacterial growth culture media with and without human cell culture media added. A human T cell proliferation assay was used as measurement for SAg activity and simultaneously we also measured the cytokine content in normal human peripheral blood leucocyte cell culture media. The results revealed that invasive and non-invasive isolates did not differ significantly in SAg activity as it is present in semipurified bacterial culture medium.

OL-064 Superantigen gene profiles and presence of exfoliative toxin genes in community-acquired meticillin-resistant Staphylococcus aureus isolated from Chinese children

OL-064 Superantigen gene profiles and presence of exfoliative toxin genes in community-acquired meticillin-resistant Staphylococcus aureus isolated from Chinese children

Superantigenic Toxin Genes Coexist with Specific Staphylococcal Cassette Chromosome mec Genes in Methicillin-Resistant Staphylococcus aureus

Staphylococcus aureus is a leading cause of human disease in the hospital setting and the community. Superantigenic toxin-producing methicillin-resistant Staphylococcus aureus (MRSA) is currently important for nosocomial infections and food-borne diseases worldwide because of its global spreading and difficulty in therapy. Superantigenic toxins can bypass normal antigen presentation and have strong T cell mitogenic activity, leading to massive release of proinflammatory cytokines and contributing to the severity of S. aureus sepsis. In this study, a total of 131 MRSA isolates from patients in the University Hospital were searched for staphylococcal cassette chromosome mec (SCCmec) genes and the staphylococcal superantigenic toxin genes by multiplex polymerase chain reactions. The MRSA isolates were classified into SCCmec type II (74.8%), type I (13.0%), type IV (3.8%), type V (2.3%), and type I and type II (3.8%). MRSA isolates (102/131) also carried a number of superantigenic toxin genes including staphylococcal enterotoxin (se) and toxic shock syndrome toxin-1 (tst-1) genes. The most frequent superantigen gene profile (55/131, 42.0%) of the MRSA isolates includes staphylococcal enterotoxin C (sec), seg, sei, staphylococcal enterotoxin-like L (sell), selm, seln, selo, and tst-1. Furthermore, SCCmec type I or type II MRSA isolates more frequently harbor sec, seg, sei, sell, selm, seln, selo, and tst-1 genes, compared to other types of MRSA. These results indicate that the selected superantigenic toxin genes are linked to SCCmec type I and type II. The coexistence of these toxins and the SCCmec genes in S. aureus may contribute to the biological fitness and pathogenicity of MRSA.

Emm gene diversity, superantigen gene profiles and presence of SlaA among clinical isolates of group A, C and G streptococci from western Norway

In order to investigate molecular characteristics of beta-hemolytic streptococcal isolates from western Norway, we analysed the entire emm gene sequences, obtained superantigen gene profiles and determined the prevalence of the gene encoding streptococcal phospholipase A2 (SlaA) of 165 non-invasive and 34 contemporary invasive group A, C and G streptococci (GAS, GCS and GGS). Among the 25 GAS and 26 GCS/GGS emm subtypes identified, only emm3.1 was significantly associated with invasive disease. M protein size variation within GAS and GCS/GGS emm types was frequently identified. Two non-invasive and one invasive GGS possessed emm genes that translated to truncated M proteins as a result of frameshift mutations. Results suggestive of recombinations between emm or emm-like gene segments were found in isolates of emm4 and stG485 types. One non-invasive GGS possessed speC, speG, speH, speI and smeZ, and another non-invasive GGS harboured SlaA. speA and SlaA were over-represented among invasive GAS, probably because they were associated with emm3. speG dys was identified in 83% of invasive and 63% of non-invasive GCS/GGS and correlated with certain emm subtypes. Our results indicate the invasive potential of isolates belonging to emm3, and show substantial emm gene diversity and possible lateral gene transfers in our streptococcal population.

Superantigen gene profiles and presence of exfoliative toxin genes in community-acquired meticillin-resistant Staphylococcus aureus isolated from Chinese children

This study aimed to evaluate the distribution of superantigen gene profiles and the presence of exfoliative toxin genes in community-acquired meticillin-resistant Staphylococcus aureus (CA-MRSA) isolated from Chinese children, and simultaneously to assess virulence gene profiles and genetic background. Of the CA-MRSA isolates, 88.9 % (88/99) harboured toxin genes, with sek as the most frequent toxin gene (62.6 %), followed by seq (61.6 %), seb (60.6 %) and sea (35.4 %). The eta gene was detected only in one ST398-IVa-spa t034 strain. The sed and etd genes were not found in any of the isolates tested. A total of 38 virulence genotypes were observed, of which the genotype seb-sek-seq (27.3 %, 24/88) comprised the majority, followed by sea-seb-sek-seq (18.2 %, 16/88). The enterotoxin gene cluster including seg-sei-sem-sen-seo-seu predominated at a rate of 15.1 %. The relationship among toxin genotypes, toxin genes encoding profiles of mobile genetic elements and genetic background was analysed. Among 66 clonal complex (CC) 59 isolates, 87.9 % (58/66) were positive for toxin genes, and 75.8 % (50/66) harboured the toxin gene combination seb-sek-seq. Among seb-sek-seq-positive CC59 strains, 42.0 % (21/50) also carried the sea gene. CC59 corresponded exclusively to accessory gene regulator 1 (agr-1). The data presented here enhance our current knowledge on the virulence determinants of CA-MRSA.

Superantigen Genes Are More Important than theemmType for the Invasiveness of Group AStreptococcusInfection

In this study, we examined the role of superantigen genes and emm genotypes of clinical Streptococcus pyogenes isolates collected in Germany between 1997 and 2003. Multiplex polymerase chain reaction for all 11 currently known superantigen genes and sequencing for emm types were used. Using a 2-step explorative data analysis procedure, we found that after combined analysis of superantigen genes and emm types, only the superantigen genes spea1-spea3, spem, and spea4 have a predictive value for invasiveness, with odds ratios of 7.992, 3.209, and 2.323, respectively. The predictive value for invasiveness of emm1 was lost after combined analysis because of the association between emm type and the highly predictive superantigen genes. On the other hand, presence of the superantigen gene ssa and of emm77 are predictors of noninvasiveness, with odds ratios of 0.370 and 0.271, respectively. These results indicate that the presence of superantigen genes is more important for the invasiveness of group A Streptococcus infection than emm type and may be the connection between the high-risk HLA type of the host and the pathogen. Furthermore, we found a very clear correlation between the presence of the genes spea1-spea3 and the presence of the gene emm1, which indicates that the relationship between emm1 and invasiveness is based on the superantigen gene profile. Our data suggest that the superantigen gene profile is of high importance for the clinical outcome of group A Streptococcus infections.

Clinical, microbiological and molecular characteristics of six cases of group A streptococcal meningitis in western Norway

Meningitis is a rare clinical manifestation of invasive group A streptococcal (iGAS) disease. Clinical, microbiological and molecular characteristics of 6 consecutive cases of GAS meningitis treated in Haukeland University Hospital in the period 2004-2009 are described. All 6 patients had a primary upper respiratory tract infection, with subsequent mastoiditis in 5, subdural effusions in 2, and cerebral abscess in 1. Five patients needed surgical treatment (myringotomy, craniotomy or mastoidectomy). All patients were treated with a beta-lactam antibiotic in combination with rifampicin. The course was complicated in all cases, and 1 patient died. Three of the bacterial isolates were of the sequence type emm1.0 and they shared the same superantigen gene profile (speA, speG, speJ, smeZ). The remaining 3 isolates belonged to sequence types emm 3.1, emm6.4 and emm12.0. Deletions in emm genes were observed. This report describes the severe and complicated course of GAS meningitis and its management, often requiring surgical intervention.