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%). 450 bp Size band in prtF1 and 350 bp 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.     

Development of an opsonophagocytic killing assay for group a streptococcus

Group A Streptococcus (GAS) or Streptococcus pyogenes is responsible for an estimated 500,000 deaths worldwide each year. Protection against GAS infection is thought to be mediated by phagocytosis, enhanced by bacteria-specific antibody. There are no licenced GAS vaccines, despite many promising candidates in preclinical and early stage clinical development, the most advanced of which are based on the GAS M-protein. Vaccine progress has been hindered, in part, by the lack of a standardised functional assay suitable for vaccine evaluation. Current assays, developed over 50 years ago, rely on non-immune human whole blood as a source of neutrophils and complement. Variations in complement and neutrophil activity between donors result in variable data that is difficult to interpret. We have developed an opsonophagocytic killing assay (OPKA) for GAS that utilises dimethylformamide (DMF)-differentiated human promyelocytic leukemia cells (HL-60) as a source of neutrophils and baby rabbit complement, thus removing the major sources of variation in current assays. We have standardised the OPKA for several clinically relevant GAS strain types (emm1, emm6 and emm12) and have shown antibody-specific killing for each emm-type using M-protein specific rabbit antisera. Specificity was demonstrated by pre-incubation of the antisera with homologous M-protein antigens that blocked antibody-specific killing. Additional qualifications of the GAS OPKA, including the assessment of the accuracy, precision, linearity and the lower limit of quantification, were also performed. This GAS OPKA assay has the potential to provide a robust and reproducible platform to accelerate GAS vaccine development.     

Diversity of emm sequence types in group A beta-haemolytic streptococci in two remote Northern Territory Indigenous communities: Implications for vaccine development

There is a high burden of disease due to group A streptococcus (GAS) in remote Northern Territory (NT) Indigenous communities. A proposed 26-valent GAS M-type vaccine covers 80–90% of pharyngeal and invasive isolates in the US. We examined the diversity and distribution of emm types in two remote Indigenous communities in the NT Top End over a 17-year period and compared them to the proposed vaccine types. Eighty emm types were identified between 1991 and 2007. Diversity in both communities was high (overall Simpson's index 0.976), but varied between communities. Prior to 2004, 71 emm types were identified and an additional 9 emm types were identified during a period of active surveillance in 2004–2005. The proposed 26-valent vaccine would be expected to cover only 20% of emm types recovered in this study. Of the 80 emm types, 16 (20%) were new sequence types identified since the last assignment of M types in 2002. The diversity of streptococcal isolates was higher than that reported from most industrialized countries, and similar to that described in several developing countries. A vaccine based on such a variable antigen is unlikely to provide effective protection in the highest risk populations.     

Characteristics of Group A Streptococcus Strains Circulating during Scarlet Fever Epidemic,Beijing, China, 2011

Scarlet fever is one of a variety of diseases caused by group A Streptococcus (GAS). During 2011, a scarlet fever epidemic characterized by peak monthly incidence rates 2.9–6.7 times higher than those in 2006–2010 occurred in Beijing, China. During the epidemic, hospital-based enhanced surveillance for scarlet fever and pharyngitis was conducted to determine characteristics of circulating GAS strains. The surveillance identified 3,359 clinical cases of scarlet fever or pharyngitis. GAS was isolated from 647 of the patients; 76.4% of the strains were type emm12, and 17.1% were emm1. Almost all isolates harbored superantigens speC and ssa. All isolates were susceptible to penicillin, and resistance rates were 96.1% to erythromycin, 93.7% to tetracycline, and 79.4% to clindamycin. Because emm12 type GAS is not the predominant type in other countries, wider surveillance for the possible spread of emm12 type GAS from China to other countries is warranted.     

Molecular characterization and antibiotic resistance of clinical Streptococcus dysgalactiae subsp. equisimilis in Beijing,China

Streptococcus dysgalactiae subsp. equisimilis (SDSE) is presently considered as a human pathogen associated with clinical infection. We characterized 56 SDSE isolates collected from two tertiary hospitals in Beijing, China. Sixteen distinct emm types/subtypes were detected, dominated by stG245.0 (32.1%), stG652.0 (10.7%), stG6.1 (10.7%) and stG485.0 (10.7%), and a novel stG840.0 variant type was identified. All isolates possessed virulence genes of sagA and scpA, and most carried slo (98.2%), ska (98.2%) and speG^dys (35.7%). By multilocus sequence typing (MLST) analysis, 17 individual sequence types (STs) were distinguished, including 7 newly-identified STs (26.8% of isolates), of which ST127 (30.4%), ST7 (12.5%) and ST44 (10.7%) dominated. Meanwhile, pulsed-field gel electrophoresis (PFGE) analysis revealed 33 pattern types (PTs), which were further combined into 16 pattern clusters (PCs), and 59.3% of isolates were distributed into 2 dominant PCs. Notably, emm types had both close relationship and consistency with STs and PFGE PCs. Furthermore, of 56 SDSE isolates, the predominant antibiotic resistances were erythromycin (71.4%), clindamycin (71.4%) and tetracycline (60.7%). Correspondingly, the prevalent resistance genes of macrolide and tetracycline were erm(B) (78.6%) and tet(M) (73.2%). In addition, multiple point mutations of parC, one of fluoroquinolone resistance genes, were observed (accounting for 75%), and were divided into 12 types, with parC 07 as the predominant type. Our data suggested the wide molecular diversity and distinctive regional features of SDSE from clinical infection in Beijing, China.     

Potential coverage of a multivalent M protein-based group A streptococcal vaccine

Background

The greatest burden of group A streptococcal (GAS) disease worldwide is due to acute rheumatic fever (ARF) and rheumatic heart disease (RHD). Safe, effective and affordable vaccines designed to prevent GAS infections that trigger ARF could reduce the overall global morbidity and mortality from RHD. The current study evaluated the potential coverage of a new 30-valent M protein-based vaccine using GAS isolates from school children in Bamako, Mali, a population at high risk for the development of RHD.

Methods

The bactericidal activity of rabbit antisera against the 30-valent vaccine was assessed using a collection of GAS isolates recovered during a study of the epidemiology of pharyngitis in Bamako.

Results

Single isolates representing 42 of 67 emm-types, accounting for 85% of the GAS infections during the study, were evaluated. All (14/14) of the vaccine emm-types in the collection were opsonized (bactericidal killing >50%) and 26/28 non-vaccine types were opsonized. Bactericidal activity was observed against 60% of the total emm-types recovered in Bamako, which accounted for 81% of all infections.

Conclusions

Multivalent vaccines comprised of N-terminal M peptides elicit bactericidal antibodies against a broad range of GAS serotypes, indicating that their efficacy may extend beyond the emm-types included in the vaccine.     

山东省2013年致猩红热及无症状携带者A群链球菌分子分型研究

目的 描述2013年山东省致猩红热及无症状携带者A群链球菌(GAS)分子分型特征,探讨各分型方法与emm型别的关联.方法 自猩红热、无症状携带者咽拭分离获得GAS 72株,利用emm、多位点序列分型(MLST)、超抗原(SAg)基因(speA、speC、speG、speH、speI、speJ、smeZ、ssa、speK、speM、speL)检测和脉冲场凝胶电泳(PFGE)对其进行分型检测.结果 72株GAS中,以emm1型(41.67%)和emm12型(56.94%)为主;共检出两种ST型别,其中ST28型31株(43.06%),ST36型41株(56.94%);ST28型与emm1相关(P<0.05),而ST36型与emm12相关(P<0.05);speA、speC、speG、speH、speI、speJ、smeZ、ssa的检出率分别为38.89%、95.83%、97.22%、38.89%、54.17%、41.67%、100.00%、100.00%,speK、speM、speL未检出;emm1型菌株倾向于含有speA、speJ,不含有speH、speI(P<0.05);emm12恰好相反,倾向于含有speH、speI,不含有speA、speJ(P<0.05);PFGE共检出20种型别.结论 2013年山东省致猩红热和无症状携带者中GAS型别分布较为单一,emm型别以emm1和emm12为主;ST型别主要为ST28和ST36,超抗原基因携带以speC、speG、smeZ、ssa为主.各分型方法间存在一定关联.     

Group A Streptococcus emm gene types in pharyngeal isolates, Ontario, Canada, 2002-2010

Group A Streptococcus (GAS) is a human-adapted pathogen that causes a variety of diseases, including pharyngitis and invasive infections. GAS strains are categorized by variation in the nucleotide sequence of the gene (emm) that encodes the M protein. To identify the emm types of GAS strains causing pharyngitis in Ontario, Canada, we sequenced the hypervariable region of the emm gene in 4,635 pharyngeal GAS isolates collected during 2002-2010. The most prevalent emm types varied little from year to year. In contrast, fine-scale geographic analysis identified inter-site variability in the most common emm types. Additionally, we observed fluctuations in yearly frequency of emm3 strains from pharyngitis patients that coincided with peaks of emm3 invasive infections. We also discovered a striking increase in frequency of emm89 strains among isolates from patients with pharyngitis and invasive disease. These findings about the epidemiology of GAS are potentially useful for vaccine research.     

Changes in antimicrobial resistance and molecular attributes of Salmonellae causing enteric fever in Kolkata,India, 2014–2018

Globally, enteric fever caused by Salmonella Typhi (S. Typhi, ST) and S. Paratyphi A (SPA) remain one of the major diseases of public health importance. In this study, a total of 457 (380 ST, 77 SPA) blood isolates were collected from three tertiary care hospitals in Kolkata during 2014–18. Additionally, 66 (3.4%) ST and 5 (0.25%) SPA were recovered from blood culture of 1962 patients attending OPD of one pediatric hospital during 2016–18. The study isolates were tested for antimicrobial resistance (AMR) profiles; AMR genes; molecular sub-types by PFGE, MLVA and CRISPR. Among the total 446 ST and 82 SPA isolates, fluoroquinolone (FQ) resistance was very common in both serovars. Ciprofloxacin resistance of 24.9% and 9.8% & ofloxacin resistance of 20.9% and 87.8% were found in ST and SPA respectively. Majority (>70%) of the isolates showed decreased susceptibility to ciprofloxacin (DCS). A single point mutation in gyrA gene (S83F) was responsible for causing DCS in 37.5% (n = 42/112) ST and 63% (n = 46/73) SPA isolates. Multidrug resistance (MDR) was found only in 3.4% ST isolates and encoded the genes bla_TEM-1, catA, sul, strA-strB, class 1 integron with dfrA7. All MDR ST (n = 15) possessed non-conjugative non-IncHI1 (180 kb) plasmid except one having conjugative IncHI1 (230 kb) plasmid and one without plasmid. The MDR genes were integrated near chromosomal cyaA gene site in ST with/without the presence of plasmid (nonIncH1). Almost 65.7% resistant ST belonged to H58 haplotype. PFGE showed clonally related isolates with 81% similarity in ST and 87% in SPA. Similarly, CRISPR typing showed less diversity among the isolates. However, the isolates (ST and SPA) were found to be more diverse by MLVA typing (D value 0.987 and 0.938). The study reports decrease in MDR and increase in FQ resistance among typhoidal Salmonella isolates over the years giving interesting information for enteric fever treatment.     

Invasive Multidrug-Resistant emm93.0 Streptococcus pyogenes Strain Harboring a Novel Genomic Island,Israel, 2017–2019

Invasive group A Streptococcus (iGAS) infections have increased in Israel since 2016 as successful lineages have emerged. We report the emergence and outbreak of a multidrug-resistant S. pyogenes emm93.0, sequence type 10, among iGAS infections in Israel since 2017. This type has been observed very rarely in other countries. During this period, emm93.0 was the cause of 116 infections in Israel and became the leading type during 2018. Most of the infections were from bacteremia (75%), and most patients were male (76%). We observed infections across Israel, mainly in adults. Of note, we observed multidrug resistance for clindamycin, tetracycline, and trimethoprim/sulfamethoxazole. Whole-genome sequencing confirmed clonality among geographically disseminated isolates. The local emm93.0 sequence type 10 clone contained a novel genomic island harboring the resistance genes lsa(E), lnu(B), and ant (6)-Ia aph(3′)-III. Further phenotypic and genomic studies are required to determine the prevalence of this resistance element in other iGAS types.

Streptococcus pyogenes is a major cause of community-acquired and nosocomial infections linked with illness and death worldwide (1,2). Group A Streptococcus (GAS) species cause a variety of infections, including pharyngitis, skin and soft tissue infections (SSTI), severe invasive infections, bacteremia, and toxic shock syndrome (3, 4). Acquisition of GAS is mainly attributed to person-to-person transmission by respiratory droplets or skin contact; in addition, inoculated food can spread the infection, resulting in outbreaks (5,6).Surveillance programs and prevention guidelines focus on systemic GAS infection, which is defined as a statutory notifiable disease in many countries (5). Active laboratory-based surveillance based on molecular characterization of invasive isolates is essential for outbreak detection and public health response. However, laboratory surveillance data indicate that most invasive cases in industrialized countries occur sporadically and are not part of outbreak clusters (5). In Israel, iGAS is a notifiable disease; all invasive isolates are analyzed at the national reference laboratory at the Ministry of Health (Jerusalem, Israel).The emm typing scheme is a primary tool for surveillance, outbreak detection, and for the study of the population structure on the basis of the sequencing of the emm gene (7). Specific emm types and M proteins (M1, M3) have been linked with invasive infections. Several studies show that emm types are significantly more diverse in developing countries than in developed countries (5).Susceptibility testing is essential for successful outbreak control. Although β-lactams are the preferred antimicrobial drug treatment for GAS infections, macrolides, lincosamides, and streptogramins are useful for treating patients with β-lactam allergy, and for overcoming treatment failure in patients treated with penicillin. Recommendations for treating severe invasive cases include adding clindamycin or linezolid, which suppress toxin production (8,9). Several reports highlight the emergence of successful clones associated with acquired antimicrobial resistance (10–12). Clindamycin resistance is rarely reported and is associated with specific lineages (13,14).We describe the epidemiology of invasive GAS (iGAS) in Israel during 2014–2019. We report the emergence and ongoing outbreak of a multidrug-resistant (MDR) S. pyogenes emm93.0 that caused 116 iGAS cases during 2016–2019 in Israel. The unique epidemiologic dynamics of the outbreak clone, as well as in-depth whole-genome sequence analysis, were the focus of our investigation.     

Molecular Characterization of Invasive Streptococcus dysgalactiae subsp. equisimilis,Japan

We collected β-hemolytic streptococci (1,611 isolates) from patients with invasive streptococcal infections in Japan during April 2010–March 2013. Streptococcus dysgalactiae subsp. equisimilis (SDSE) was most common (n = 693); 99% of patients with SDSE infections were elderly (mean age 75 years, SD ±15 years). We aimed to clarify molecular and epidemiologic characteristics of SDSE isolates and features of patient infections. Bacteremia with no identified focus of origin and cellulitis were the most prevalent manifestations; otherwise, clinical manifestations resembled those of S. pyogenes infections. Clinical manifestations also differed by patient’s age. SDSE isolates were classified into 34 emm types; stG6792 was most prevalent (27.1%), followed by stG485 and stG245. Mortality rates did not differ according to emm types. Multilocus sequence typing identified 46 sequence types and 12 novel types. Types possessing macrolide- and quinolone-resistance genes were 18.4% and 2.6%, respectively; none showed β-lactam resistance. Among aging populations, invasive SDSE infections are an increasing risk.     

Molecular characterization of Latin American invasive Streptococcus pneumoniae serotype 19A isolates

Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide, especially among children and the elderly. S. pneumoniae serotype 19A has emerged as a major cause of invasive disease in many countries, regardless of whether pneumococcal conjugate vaccines are used. The aim of this study was molecular characterization of invasive S. pneumoniae serotype 19A isolates recovered between 2000 and 2015 from 13 National Laboratories through the laboratory-based surveillance of invasive S. pneumoniae program SIREVA II in Latin American countries. The isolates were submitted with antimicrobial susceptibility tests and were genotyped by a combination of pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Of the 185 isolates assayed, notable rates of resistance to penicillin (MIC ≥ 0.125 µg/mL; 68.6%), tetracycline (63.7%), trimethoprim-sulfamethoxazole (63.2%), and erythromycin (43.2%) were observed, while 44.3% of isolates were multidrug resistant. The most frequently observed sequence types (ST) were ST320 (32.4%), ST199 (14.1%), ST172 (10.8%) and ST5204 (7.1%). The distribution of STs indicated regional differences in the epidemiology of the clonal groups. The present study showed a diverse genetic background of the pneumococcal population in Latin American countries. Continuous surveillance of the pneumococcal serotype 19A population in the region will be necessary to obtain information about geographical differences and changes in the spread and the establishment of particular clones.     

Genotypic diversity of multidrug-, quinolone- and extensively drug-resistant Mycobacterium tuberculosis isolates in Thailand

Drug-resistant tuberculosis (TB), which includes multidrug-resistant (MDR-TB), quinolone-resistant (QR-TB) and extensively drug-resistant tuberculosis (XDR-TB), is a serious threat to TB control. We aimed to characterize the genotypic diversity of drug-resistant TB clinical isolates collected in Thailand to establish whether the emergence of drug-resistant TB is attributable to transmitted resistance or acquired resistance. We constructed the first molecular phylogeny of MDR-TB (n = 95), QR-TB (n = 69) and XDR-TB (n = 28) in Thailand based on spoligotyping and proposed 24-locus multilocus variable-number of tandem repeat analysis (MLVA). Clustering analysis was performed using the unweighted pair group method with arithmetic mean. Spoligotyping identified the Beijing strain (SIT1) as the most predominant genotype (n = 139; 72.4%). The discriminatory power of 0.9235 Hunter–Gaston Discriminatory Index (HGDI) with the 15-locus variable-number tandem repeats of mycobacterial interspersed repetitive units typing was improved to a 0.9574 HGDI with proposed 24-locus MLVA, thereby resulting in the subdivision of a large cluster of Beijing strains (SIT1) into 17 subclusters. We identified the spread of drug-resistant TB clones caused by three different MLVA types in the Beijing strain (SIT1) and a specific clone of XDR-TB caused by a rare genotype, the Manu-ancestor strain (SIT523). Overall, 49.5% of all isolates were clustered. These findings suggest that a remarkable transmission of drug-resistant TB occurred in Thailand. The remaining 50% of drug-resistant TB isolates were unique genotypes, which may have arisen from the individual acquisition of drug resistance. Our results suggest that transmitted and acquired resistance have played an equal role in the emergence of drug-resistant TB. Further characterization of whole genome sequences of clonal strains could help to elucidate the mycobacterial genetic factors relevant for drug resistance, transmissibility and virulence.     

Evaluation of a new trilocus sequence-based multiplex-PCR to detect major Acinetobacter baumannii clonal complexes circulating in Brazil

Dissemination of carbapenem-resistant Acinetobacter baumannii (CRAB) is mainly due to the spread of clonal lineages, particularly those included into the clonal complexes (CC) CC1, CC2, CC15, CC25, and CC79. We evaluated the usefulness of a recently modified PCR-based trilocus sequence-based typing (m3LST) in comparison with the standard multilocus sequence typing (MSLT) of 7 housekeeping genes as per the Institute Pasteur Scheme to assign the clonal complexes in CRAB. A collection of 78 CRAB isolated from 67 different Brazilian health institutions was submitted to both methodologies, and concordance rate was calculated. The collection studied included mainly isolates belonging to endemic Brazilian Clonal Complexes (CC1, CC15, CC25 and CC79, n = 72, 92.3%) but also singletons sequence types (ST) with low prevalence in the country (ST107, ST113, ST188, ST317, ST584, ST733, n = 6; 7.7%). The m3LST correctly assigned all the isolates into the main CC responsible for the CRAB dissemination in Brazil. All the singletons ST were not misidentified as prevalent lineages. The PCR-based m3LST is a powerful tool to investigate molecular epidemiology of A. baumannii representative of prevalent Brazilian clonal complexes 1, 15, 25 and 79.     

Multiple-locus variable number tandem repeat analysis of Neisseria gonorrhoeae isolates in Russia

In the present study, a multiple-locus variable number tandem repeat analysis (MLVA) was used to assess the molecular epidemiology of Neisseria gonorrhoeae clinical isolates originating from different regions of Russia. MLVA, based on seven loci, was performed on 218 isolates that were previously tested for susceptibility to penicillin, tetracycline and ciprofloxacin and for the presence of certain genetic determinants of drug resistance. In total, 83 MLVA types were identified, indicating that MLVA is a highly discriminatory technique with a Hunter–Gaston discriminatory index of 0.963 (95% CI, 0.950–0.977). MLVA type 16 was shown to be the most prevalent type and is undoubtedly associated with a multidrug resistant phenotype. The spread of MLVA type 16 from Moscow to Irkutsk suggests that this type has a highly successful transmission rate. Hierarchical cluster analysis of the MLVA profiles classified 208 isolates (95%) into six large groups (containing more than 10 isolates). Clusters differed in geographical characteristics and susceptibility profiles. MLVA cluster A comprised in total 34 isolates and was unambiguously associated with multidrug resistance. Most isolates in cluster A carried mutations in penA, ponA, rpsJ, mtrR, gyrA, and parC genes. MLVA cluster D was associated with resistance to penicillin and with mutations in ponA and rpsJ genes and the presence of plasmid-borne bla_(TEM-1) gene. MLVA clusters B, C and E were associated with susceptibility to ciprofloxacin and had a lack of mutations in ponA, rpsJ, gyrA, and parC genes. We conclude that MLVA will be a useful tool for N. gonorrhoeae epidemiological studies.     

Phylogenetic analysis of carbapenem-resistant Acinetobacter baumannii isolated from different sources using Multilocus Sequence Typing Scheme

The emergence and worldwide distribution of carbapenem-resistant Acinetobacter baumannii strains has become a major public health threat. The objective of this study was to investigate the clonal relatedness of A. baumannii isolates collected from clinical and extra-hospital environments in Mthatha, South Africa. Forty carbapenem-resistant isolates comprising of clinical (20) and extra-hospital (20) were identified and tested for antimicrobial susceptibility. Detection of carbapenemase encoding genes was performed by Real-time PCR. The clonal relationship of clinical isolates relative to extra-hospital isolates was determined via multilocus sequence typing (MLST). All isolates (clinical and extra-hospital) were resistant to most common antibiotics including carbapenems (imipenem; MIC ≥32 μg/mL and meropenem; MIC ≥32 μg/mL) with the only exception being amikacin (with 3 isolates susceptible), tigecycline (14 isolates susceptible) and colistin (all isolates susceptible). The bla OXA-23-like and the intrinsic bla OXA-51 -like genes were detected in all the isolates tested. The bla OXA-58-like and bla IMP-type genes were detected in 2 clinical isolates whilst the bla OXA-24-like, bla VIM-type, bla NDM-1, bla SIM, and bla AmpC were not detected. The bla OXA-24-like, bla OXA-58-like, bla IMP-type, bla VIM-type, bla NDM-1, bla SIM, and bla AmpC were negative in the extra-hospital isolates. Co-occurrence of bla OXA-23 -like, bla OXA-58-like and bla IMP-type was observed in 2 clinical isolates. The MLST performed on 33 isolates identified 5 existing sequence types (ST) (ST1, ST2, ST25, ST85 and ST215) in clinical isolates and 2 existing STs (ST1 and ST2) in extra-hospital isolates. The most dominant ST was ST2 accounting for 68.8% of the clinical isolates and 82.4% of the extra-hospital isolates. The study demonstrated high prevalence and potential clonal spread of globally-disseminated clonal complex 2 carrying bla OXA-23-like within our local settings. However, ST25 might be an emerging lineage carrying the bla OXA-23-like . Continuous monitoring is important in limiting the spread of these strains in other healthcare settings and the community.     

Whole genome characterization of Streptococcus pneumoniae from respiratory and blood cultures collected from Canadian hospitals before and after PCV-13 implementation in Canada: Focus on serotypes 22F and 33F from CANWARD 2007–2018

The population of pneumococci circulating in Canada is constantly shifting under the pressures of antimicrobial and conjugate vaccine use. A new 15-valent pneumococcal conjugate vaccine (PCV), containing PCV-13 serotypes plus additional serotypes 22F and 33F, is currently undergoing clinical trials. The purpose of this study was to utilize whole genome sequencing to characterize invasive and respiratory Streptococcus pneumoniae isolates collected from Canadian hospitals pre- (2007–2011) and post-PCV-13 implementation (2012–2018) in Canada, particularly serotypes 22F and 33F. Isolates were obtained from the CANWARD 2007 to 2018 study. Overall, 597 S. pneumoniae isolates were sequenced using the Illumina MiSeq platform: 180 (101 respiratory, 79 blood) isolates of serotype 22F, 74 (41 respiratory, 33 blood) isolates of serotype 33F and 343 isolates randomly selected to broadly encompass pneumococci in Canada. Genomes were clustered using PopPUNK v2.0.2 and assigned to a Global Pneumococcal Sequencing Cluster (GPSC) and MLST sequence type (ST), and visualized using Cytoscape v3.8.0. Acquired resistance genes were identified using ResFinder 2.1, and genes with chromosomal mutations conferring resistance were extracted and compared to standard reference genome R6. PopPUNK clustering suggests that a clone of S. pneumoniae serotype 22F/ST433/GPSC19 demonstrating mefA-mediated macrolide resistance is emerging in Canada post-PCV-13 introduction, collected from both invasive and respiratory sources. Similarly, there is evidence to support a post-PCV-13 shift towards macrolide- and trimethoprim/sulfamethoxazole-resistant serotype 33F/ST100/GPSC3, including a cluster associated with invasive isolates. While some lineages containing vaccine serotypes were predominantly identified pre-PCV-13 implementation (serotype 5/GPSC8, serotype 7F/GPSC15), others (serotype 19A/GPSC1 and 4, serotype 3/GPSC12) continue to maintain a significant presence over time despite inclusion in PCV-13. Further genomic surveillance is necessary to determine additional trends over time in these upcoming vaccine serotypes, as well as the overall pneumococcal population in Canada.     

Nursing home outbreak of invasive group a streptococcal infections caused by 2 distinct strains.

OBJECTIVE: To identify factors contributing to a cluster of deaths from invasive group A streptococcus (GAS) infection in a nursing home facility and to prevent additional cases. DESIGN: Outbreak investigation. SETTING: A 146-bed nursing home facility in northern Nevada. METHODS: We defined a case as the isolation of GAS from a normally sterile site in a resident of nursing home A. To identify case patients, we reviewed resident records from nursing home A, the local hospital, and the hospital laboratory. We obtained oropharyngeal and skin lesion swabs from staff and residents to assess GAS colonization and performed emm typing on available isolates. To identify potential risk factors for transmission, we performed a cohort study and investigated concurrent illness among residents and surveyed staff regarding infection control practices. RESULTS: Six residents met the case patient definition; 3 (50%) of them died. Among invasive GAS isolates available for analysis, 2 distinct strains were identified: emm11 (3 isolates) and emm89 (2 isolates). The rate of GAS carriage was 6% among residents and 4% among staff; carriage isolates were emm89 (8 isolates), emm11 (2 isolates), and emm1 (1 isolate). Concurrently, 35 (24%) of the residents developed a respiratory illness of unknown etiology; 41% of these persons died. Twenty-one (30%) of the surveyed employees did not always wash their hands before patient contacts, and 27 (38%) did not always wash their hands between patient contacts. CONCLUSIONS: Concurrent respiratory illness likely contributed to an outbreak of invasive GAS infection from 2 strains in a highly susceptible population. This outbreak highlights the importance of appropriate infection control measures, including respiratory hygiene practices, in nursing home facilities.     

Clonal change of carbapenem-resistant Acinetobacter baumannii isolates in a Korean hospital

The expansion of specific carbapenem-resistant Acinetobacter baumannii (CRAB) clones is a global concern due to its therapeutic difficulty and epidemicity. To understand the prevalence of CRAB isolates in a Korean hospital, we investigated the epidemiological characteristics of 96 CRAB isolates between 2016 and 2018, including the sequence types (STs), antimicrobial susceptibility, and genetic background of resistance to carbapenems and aminoglycosides. Six STs were identified using the Oxford multilocus sequence typing scheme; ST191 (n = 8), ST208 (n = 12), ST229 (n = 11), and ST369 (n = 21) were previously identified clones in the study hospital, whereas gpi variants of ST208, ST451 (n = 34) and ST784 (n = 10), were emerging clones. ST208 isolates exhibited higher resistance rates to minocycline than other ST isolates, whereas ST369 isolates exhibited lower resistance rates to aminoglycosides and trimethoprim/sulfamethoxazole than other ST isolates. All CRAB isolates previously isolated in the study hospital carried ISAbaI-bla_OXA-23 for carbapenem resistance, but 10 ST229 isolates carried only ISAbaI-bla_OXA-51. The carriage of armA was lower in ST369 isolates (38%) than in other ST isolates (≥83%). The frequency and diversity of aminoglycoside-modifying enzyme genes were decreased among the CRAB isolates between 2016 and 2018 compared with CRAB isolates between 2013 and 2015 at the study hospital. In conclusion, clonal complex 208 CRAB isolates are predominant in the study hospital. This study demonstrates the evolutionary change of CRAB isolates in the study hospital in relation to the emergence of new STs and selection of resistant genes.     

北京地区2011－2014年致儿童猩红热A组链球菌emm基因型别变化特征分析

目的 了解北京地区致儿童猩红热A组链球菌(GAS)emm基因型别分布,并探讨其型别变化与猩红热病例的关系。方法 收集2011－2014年5－7月北京市36家医院临床诊断为猩红热病例咽拭子GAS分离菌株,采用PCR联合测序方法对分离菌株进行emm分型,选取中国疾病监测信息报告管理系统中北京市猩红热病例报告数据进行分析。结果 共采集咽拭子标本2 161份,分离GAS菌株762株,阳性率为35.3%(762/2 161)。检测到7个emm型别,其中emm12型占69.4%(529/762),emm1型占29.8%(227/762),其他5种emm型别(11、22、75、89、128)占0.8%(6/762)。分析各年份emm型别构成比:2011年emm12型82.2%(295/359),emm1型16.7%(60/359),其他型(11、22、89)1.1%(4/359);2012年emm12型77.3%(123/163),emm1型23.9%(39/163),emm75型0.6%(1/163);2013年emm12型50.7%(38/75),emm1型49.3%(37/75);2014年emm12型44.2%(73/165),emm1型55.2%(91/165),emm128型0.6%(1/165)。不同年份emm型别构成比变化的差异有统计学意义(P<0.001)。2011、2012年emm12为主导菌型,2014年主导型别转换为emm1型。2011－2014年报告猩红热病例分别为6 152、2 908、2 048和3 918例,以2011年报告例数最多,2014年比2013年报告病例数增加91.31%。2011年GAS分离株emm型别存在年龄差异,其中1～5岁组分离的emm12型高于>5岁组(P<0.05)。2011、2013年分离GAS菌株存在地区差异,2011年emm1型分离株为城区高于郊区(P<0.05),2013年emm1型为郊区高于城区(P<0.05)。结论 2011－2014年北京地区GAS菌株优势菌型为emm12和emm1型;不同年份emm12、emm1变换成为主导菌型;emm主导菌型变化与猩红热病例变化趋势有关。