Usefulness of the restriction-modification test plus staphylococcal cassette chromosome mec types and Panton-Valentine leukocidin encoding phages to identify Staphylococcus aureus methicillin-resistant clones

We studied the usefulness of the restriction-modification (RM) test, staphylococcal cassette chromosome (SCC) mec types, and Panton-Valentine leukocidin (PVL)-encoding phages to identify Staphylococcus aureus methicillin-resistant lineages and to differentiate clones that belong to the same lineage. A total of 108 methicillin-resistant S. aureus (MRSA) strains were characterized by pulsed-field gel electrophoresis (PFGE)--multi-locus sequence typing (MLST)--spa-typing. The RM correctly identified the lineages CC5, CC8, CC30, and CC398, but not CC25 and CC72. The SCCmec and RM combined analysis allowed differentiation between MLST types within the same lineage. Only 5 MRSA strains were PVL-positive. Four PVL-positive USA300 isolates carried elongated-head type PVL-encoding phages, while the sequence type (ST)-30 strain carried an icosahedral-head phage. The combination of the RM test method, SCCmec types, and PVL phage identification could be useful for MRSA typing purposes.     

Molecular characterization of vancomycin-resistant Enterococcus spp. clinical isolates from Hungary and Serbia

The objectives of this work were to collect and characterize vancomycin-resistant Enterococcus faecium (VREF) clinical isolates from Hungary and Serbia and to analyse their genetic relatedness. VREF isolates were initially typed by PFGE. A selection of VREF isolates representing all participating hospitals was further examined by multiple-locus variable-number tandem repeat analysis (MLVA) and multilocus sequence typing (MLST). VanB VREF isolates (n=18) recovered from blood, urine and faecal cultures at a Budapest hospital between August 2003 and December 2004 were molecularly characterized. Macrorestriction analysis of the isolates revealed their monoclonal relatedness. A cluster of infections caused by 2 distinct VanA VREF clones recovered from 6 departments was identified in a Belgrade hospital in Serbia. The vanA resistance determinant was transferable by in vitro conjugation experiments. We also identified 2 vanA-positive E. gallinarum blood culture isolates in this Belgrade hospital. Molecular typing of representative VREF isolates from Hungary and Serbia by MLVA and MLST revealed that all tested isolates belonged to MLST complex CC17 and the corresponding MLVA cluster 1. Our results extend the documented occurrence of CC17 to a new region in Europe.     

Genetic characterization of Staphylococcus aureus isolates carrying Panton-Valentine leukocidin genes in Bangladesh

To assess the spread and genetic characteristics of Panton-Valentine leukocidin (PVL) gene-carrying Staphylococcus aureus in Bangladesh, we investigated 59 strains (49 isolates from clinical specimens and 10 isolates colonized in the nasal cavities of medical staff), including 26 methicillin-resistant S. aureus (MRSA) strains. The PVL gene was detected only in methicillin-susceptible S. aureus (MSSA) strains (7 clinical strains and 2 colonizing strains). PVL gene-positive MSSA strains were found to belong to coagulase serotypes III or VI and were classified into sequence types ST88 (CC88), ST772, and ST573 (CC1) by multilocus sequence typing, and agr types 2 or 3. These types were different from those determined for MRSA (coagulase serotypes I and IV, ST240 and ST361, and agr type 1). PVL gene-positive MSSA possessed a larger number of virulence factor genes than MRSA, although they were susceptible to more antimicrobials. These findings suggest that the PVL gene is distributed to limited populations of S. aureus clones with specific genetic traits that are distinct from MRSA in Bangladesh, but genetically close to CA-MRSA clones in the CC1 lineage reported in the United States and European countries.     

Analysis of three phenotyping methods and pulsed-field gel electrophoresis for differentiation of methicillin resistant Staphylococcus aureus isolated from two hospitals in Thailand

Methicillin resistant Staphylococcus aureus (MRSA) is an important hospital and community-acquired pathogen. Rapid and reliable epidemiologic typing is necessary for controlling the spread of MRSA outbreak. The objective of this study was to compare the phenotyping with the genotyping method to differentiate MRSA isolates obtained from the two hospitals in Thailand (central and northeastern). Seventy-four MRSA isolates were randomly collected and confirmed by the presence of mecA gene. Antibiogram, phage typing and enterotoxin production were used for the phenotyping analysis. Pulsed-field gel electrophoresis (PFGE) with Smal digestion of chromosomal DNA was used for the genotyping analysis. We found 17 distinct profiles by the 3 phenotypic typing methods and 18 PFGE types designated as 5 major types (A-E) and 13 subtypes. The most frequent PFGE types and their related subtypes found in both hospitals were A and C, comprising 54 and 27%, respectively. The antibiogram could differentiate 6 different types. All isolates were resistant to the majority of antimicrobial agents tested, but were susceptible to vancomycin and fosfomycin. Ten (13.5%) MRSA isolates produced enterotoxin A. Nontypable phage and phage type 77 were found predominantly in MRSA isolated from the northeast and central hospital, respectively. A significant correlation was found between the phenotyping and the genotyping methods and there was a good correlation between antibiogram and PFGE. Antibiogram typing alone can be used as a useful epidemiological marker for practical purposes. PFGE types A and C were the common endemic MRSA clones in both hospitals in Thailand.     

四川部分地区禽源空肠弯曲菌MLST分型及遗传进化分析

目的 为了解四川部分地区分离到的48株禽源空肠弯曲菌间的分子特征,以对来源不同的空肠弯曲菌进行分子分型及遗传进化分析。方法 本研究参照MLST数据库,选取了空肠弯曲菌MLST分型7个管家基因aspA, glnA, glyA, gltA, tkt, pgm和uncA作为目的基因分别扩增及测序,并将所得序列经比对后进行遗传进化分析。结果 48株分离株中共含24种不同的序列型分属于9种不同的克隆系及11种独特型,其中22株分离株含有新序列型,占分离株的45.83%(22/48);9种不同的克隆系中以CC-21、CC-353、CC-464克隆系所含菌株相对较多,而克隆系CC-45、CC-48最少,均为1株分离株;11种独特型共存于21株分离株中,其中以ST-8675序列型最多,为8株。遗传进化树显示,属同一克隆系的不同型别间的分离株亲缘关系较近,如同属于CC-21克隆系的ST-298与ST-760型别分离株处于同一小分支、亲缘关系较近,但不同克隆系的分离株间遗传进化关系较远,鹌鹑源分离株(V13-20)单独处于一大分支与其余禽源分离株间的亲缘关系最远。总体而言,不同地区、宿主来源的分离株呈现遗传多样性。结论 本研究中分离株间具有基因多样性,MLST分型为了解该地区空肠弯曲菌的遗传特性和分布特点提供了参考依据。     

The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA)

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of hospital-acquired infections that are becoming increasingly difficult to combat because of emerging resistance to all current antibiotic classes. The evolutionary origins of MRSA are poorly understood, no rational nomenclature exists, and there is no consensus on the number of major MRSA clones or the relatedness of clones described from different countries. We resolve all of these issues and provide a more thorough and precise analysis of the evolution of MRSA clones than has previously been possible. Using multilocus sequence typing and an algorithm, BURST, we analyzed an international collection of 912 MRSA and methicillin-susceptible S. aureus (MSSA) isolates. We identified 11 major MRSA clones within five groups of related genotypes. The putative ancestral genotype of each group and the most parsimonious patterns of descent of isolates from each ancestor were inferred by using BURST, which, together with analysis of the methicillin resistance genes, established the likely evolutionary origins of each major MRSA clone, the genotype of the original MRSA clone and its MSSA progenitor, and the extent of acquisition and horizontal movement of the methicillin resistance genes. Major MRSA clones have arisen repeatedly from successful epidemic MSSA strains, and isolates with decreased susceptibility to vancomycin, the antibiotic of last resort, are arising from some of these major MRSA clones, highlighting a depressing progression of increasing drug resistance within a small number of ecologically successful S. aureus genotypes.     

The spread of a mupirocin-resistant/methicillin-resistant Staphylococcus aureus clone in Kuwait hospitals

High-level mupirocin- and methicillin-resistant Staphylococcus aureus (MRSA) isolated from five hospitals in Kuwait were studied by pulsed-field gel electrophoresis (PFGE) to determine their relatedness to one another and to high-level mupirocin-resistant MRSA isolated previously in a Burns Unit. The genetic location of mupirocin resistance determinant was also determined. All of the isolates were resistant to gentamicin, kanamycin, streptomycin, tetracycline and cadmium, and contained plasmids of 38, 26 and 2.8 kb. Two isolates contained additional 4.4-kb plasmids. Transfer experiments demonstrated that the 38-kb plasmid encoded high-level mupirocin resistance and the 4.4-kb plasmid encoded chloramphenicol resistance. PFGE typing of representative isolates from the five hospitals demonstrated that the majority of them had identical or closely related pulsed-field patterns suggesting that they had a common origin. However, they differed from high-level mupirocin-resistant MRSA isolated previously in the Burns Unit in their resistance and pulsed-field patterns. Whereas the majority of the previous isolates were susceptible to ciprofloxacin and resistant to trimethoprim and chloramphenicol, the majority of the current isolates were susceptible to trimethoprim and chloramphenicol, and resistant to ciprofloxacin. Only one of the current isolates had identical pulsed-field pattern to the majority of isolates obtained previously in the Burns Unit. The results suggested that a previously dominant clone of high-level mupirocin-resistant MRSA has been replaced in the Burns Unit by a new clone, which also spread in four other hospitals.     

What is community-associated methicillin-resistant Staphylococcus aureus?

BACKGROUND: A community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infection has been defined as an MRSA infection in a patient who lacks specific risk factors for healthcare exposure. We sought to determine whether the absence or presence of these risk factors still predicts the phenotypic or genotypic characteristics of MRSA strains. METHODS: All clinical MRSA isolates were prospectively collected at the University of Chicago Hospitals from July 2004 through June 2005. Patients were interviewed and/or their medical records were reviewed. Isolates underwent genotyping and susceptibility testing. Data on patients and isolates were stratified in accordance with 8 frequently cited criteria for the identification of CA-MRSA and compared for concordance. RESULTS: Among 616 unique patients from whom MRSA isolates were recovered, 404 (65.6%) had risk factors for healthcare exposure. Of the 404 isolates recovered from these patients, 166 (41.1%) were clindamycin susceptible, 190 (47.0%) carried staphylococcal cassette chromosome mec (SCCmec) type IV, 145 (35.9%) carried the Panton-Valentine leukocidin genes (PVL+), and 162 (40.1%) were identified as sequence type (ST) 8 by multilocus sequence typing (MLST), all of which are characteristics commonly attributed to CA-MRSA strains. CONCLUSIONS: Association with the healthcare environment now has little predictive value for distinguishing patients with infection due to multidrug resistant MRSA isolates from those infected by CA-MRSA isolates, that is, isolates that are clindamycin-susceptible, PVL+, ST8, and/or contain SCCmec type IV. Defining CA-MRSA by the absence of risk factors for healthcare exposure greatly underestimates the burden of epidemic CA-MRSA disease.     

Concurrent Epidemics of Skin and Soft Tissue Infection and Bloodstream Infection Due to Community-Associated Methicillin-Resistant Staphylococcus aureus

Background.?Since its emergence in 2000, epidemic spread of the methicillin-resistant Staphylococcus aureus (MRSA) clone USA300 has led to a high burden of skin and soft tissue infections (SSTIs) in the United States, yet its impact on MRSA bloodstream infections (BSIs) is poorly characterized. Methods.?To assess clonality of the MRSA isolates causing SSTI and BSI during the epidemic period, a stratified, random sample of 1350 unique infection isolates (from a total of 7252) recovered at the Community Health Network of San Francisco from 2000 to 2008 were selected for genotyping. Risk factors and outcomes for 549 BSI cases caused by the USA300 epidemic clone and non-USA300 MRSA clones were assessed by retrospective review of patient medical records. Results.?From 2000 to 2008, secular trends of USA300 SSTI and USA300 BSI were strongly correlated (Pearson r?=?0.953). USA300 accounted for 55% (304/549) of BSIs as it was the predominant MRSA clone that caused community-associated (115/160), healthcare-associated community-onset (125/207), and hospital-onset (64/182) BSIs. Length of hospitalization after BSI diagnosis and mortality rates for USA300 and non-USA300 were similar. Two independent risk factors for USA300 BSI were identified: concurrent SSTI (adjusted relative risk, 1.4 [95% confidence interval {CI}, 1.2-1.6]) and anti-MRSA antimicrobial use in the preceding 30 days (0.7 [95% CI, .6-.8]). Isolates from concurrent SSTI were indistinguishable genotypically from the USA300 isolates that caused BSI. Conclusions.?USA300 SSTIs serve as a source for BSI. Strategies to control the USA300 SSTI epidemic may lessen the severity of the concurrent USA300 BSI epidemic.     

Molecular fingerprinting of mupirocin-resistant methicillin-resistant Staphylococcus aureus from a burn unit.

OBJECTIVES: To characterize mupirocin-resistant methicillin-resistant Staphylococcus aureus (MRSA) isolated from patients in a burn unit by pulsed-field gel electrophoresis and plasmid contents. METHODS: A total of 53 methicillin-resistant S. aureus, consisting of 48 mupirocin-resistant and 5 mupirocin-susceptible MRSA were compared by plasmid content and pulsed-field gel electrophoresis of Sma I digested genomic DNA. RESULTS: Of the 48 mupirocin-resistant isolates, 39 expressed high-level, and 9 expressed low-level mupirocin resistance. Plasmids were detected in all of the 53 isolates; however, only the high-level mupirocin-resistant isolates contained a 38 kb-conjugative plasmid that encoded high-level mupirocin resistance. Pulsed-field gel electrophoresis divided the isolates into four patterns designated types I to IV. Forty-three isolates consisting of 34 high-level, 5 low-level mupirocin-resistant and 4 mupirocin-susceptible isolates defined the type-I pattern. Eight isolates, five high-level and three low-level mupirocin-resistant isolates had the type-II pulsed-field pattern. The type-III and type-IV pulsed-field patterns consisted of a single isolate each. The type-I and type-II pulsed-field patterns were related and only differed by four Sma I bands. CONCLUSIONS: Results of typing the mupirocin-resistant MRSA from the burn unit with pulsed-field gel electrophoresis indicated that closely related MRSA clones previously circulating in the unit had acquired a high-level mupirocin-resistant plasmid, and spread aided by mupirocin use.     

In vivo stability of heterogeneous expression classes in clinical isolates of methicillin-resistant staphylococci

To define the stability of methicillin-resistant Staphylococcus aureus (MRSA) in vivo, 22 isolates collected at one New York institution in 1989 and 1990 were studied. All 22 belonged to one of two distinct methicillin-resistant phenotypes (class 3 or 2), which were precisely identified as belonging to two distinct genotypes. Genotypic classification was based on restriction analysis of chromosomal DNA with EcoRI and HindIII and Southern analysis of ClaI digests using two DNA probes. One was specific for the mec gene; the other was specific for transposon Tn554. The findings suggest that the MRSA isolates studied were representative of two genetically distinct MRSA "clones," each with a unique strain-specific methicillin-resistant phenotype that is stable under the conditions of invasive disease, carriage, and spread from patient to patient.     

Discriminatory powers of molecular typing techniques for methicillin-resistant Staphylococcus aureus in a university hospital, Thailand

Discriminatory powers of various molecular techniques were evaluated for typing of methicillin-resistant Staphylococcus aureus (MRSA) isolated in Siriraj Hospital, Bangkok, Thailand. Thirty MRSA isolates were randomly selected in this study. They were characterized by pulsed-field gel electrophoresis, Clal-mecA and Clal-Tn554 polymorphisms, ribotyping, and PCR-based methods including SCCmec typing, spa and coa gene polymorphism, and repeat units in hypervariable region downstream of mecA. Individual molecular typing technique distinguished those MRSA isolates into 2 to 5 types. Eleven genetic backgrounds of MRSA isolates were elucidated by combination of typing methods with trimethoprim/sulfamethoxazole (TMP/SXT) susceptibility. Combination of all typing methods including TMP/SXT susceptibility yielded a discriminatory index of 0.94. Combination of PCR-based methods and TMP/SXT susceptibility, with the discriminatory index of 0.89, is a practical typing approach suitable for rapid epidemiological investigation of MRSA isolates in a hospital setting.     

Discriminatory Indices of Typing Methods for Epidemiologic Analysis of Contemporary Staphylococcus aureus Strains

Historically, a number of typing methods have been evaluated for Staphylococcus aureus strain characterization. The emergence of contemporary strains of community-associated S. aureus, and the ensuing epidemic with a predominant strain type (USA300), necessitates re-evaluation of the discriminatory power of these typing methods for discerning molecular epidemiology and transmission dynamics, essential to investigations of hospital and community outbreaks. We compared the discriminatory index of 5 typing methods for contemporary S. aureus strain characterization.Children presenting to St. Louis Children''s Hospital and community pediatric practices in St. Louis, Missouri (MO), with community-associated S. aureus infections were enrolled. Repetitive sequence-based PCR (repPCR), pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), staphylococcal protein A (spa), and staphylococcal cassette chromosome (SCC) mec typing were performed on 200 S. aureus isolates. The discriminatory index of each method was calculated using the standard formula for this metric, where a value of 1 is highly discriminatory and a value of 0 is not discriminatory.Overall, we identified 26 distinct strain types by repPCR, 17 strain types by PFGE, 30 strain types by MLST, 68 strain types by spa typing, and 5 strain types by SCCmec typing. RepPCR had the highest discriminatory index (D) of all methods (D = 0.88), followed by spa typing (D = 0.87), MLST (D = 0.84), PFGE (D = 0.76), and SCCmec typing (D = 0.60). The method with the highest D among MRSA isolates was repPCR (D = 0.64) followed by spa typing (D = 0.45) and MLST (D = 0.44). The method with the highest D among MSSA isolates was spa typing (D = 0.98), followed by MLST (D = 0.93), repPCR (D = 0.92), and PFGE (D = 0.89). Among isolates designated USA300 by PFGE, repPCR was most discriminatory, with 10 distinct strain types identified (D = 0.63). We identified 45 MRSA isolates which were classified as identical by PFGE, MLST, spa typing, and SCCmec typing (USA300, ST8, t008, SCCmec IV, respectively); within this collection, there were 5 distinct strain types identified by repPCR.The typing methods yielded comparable discriminatory power for S. aureus characterization overall; when discriminating among USA300 isolates, repPCR retained the highest discriminatory power. This property is advantageous for investigations conducted in the era of contemporary S. aureus infections.     

Molecular tracing of the emergence, adaptation, and transmission of hospital-associated methicillin-resistant Staphylococcus aureus

Hospital-associated infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are a global health burden dominated by a small number of bacterial clones. The pandemic EMRSA-16 clone (ST36-II) has been widespread in UK hospitals for 20 y, but its evolutionary origin and the molecular basis for its hospital association are unclear. We carried out a Bayesian phylogenetic reconstruction on the basis of the genome sequences of 87 S. aureus isolates including 60 EMRSA-16 and 27 additional clonal complex 30 (CC30) isolates, collected from patients in three continents over a 53-y period. The three major pandemic clones to originate from the CC30 lineage, including phage type 80/81, Southwest Pacific, and EMRSA-16, shared a most recent common ancestor that existed over 100 y ago, whereas the hospital-associated EMRSA-16 clone is estimated to have emerged about 35 y ago. Our CC30 genome-wide analysis revealed striking molecular correlates of hospital- or community-associated pandemics represented by mobile genetic elements and nonsynonymous mutations affecting antibiotic resistance and virulence. Importantly, phylogeographic analysis indicates that EMRSA-16 spread within the United Kingdom by transmission from hospitals in large population centers in London and Glasgow to regional health-care settings, implicating patient referrals as an important cause of nationwide transmission. Taken together, the high-resolution phylogenomic approach used resulted in a unique understanding of the emergence and transmission of a major MRSA clone and provided molecular correlates of its hospital adaptation. Similar approaches for hospital-associated clones of other bacterial pathogens may inform appropriate measures for controlling their intra- and interhospital spread.     

Molecular epidemiology of methicillin-resistant Staphylococcus aureus isolates from regional hospitals in Trinidad and Tobago.

OBJECTIVES: Methicillin-resistant Staphylococcus aureus (MRSA), first reported in a British hospital in the early 1960s, has now reached global proportions. Geographic spread of one or several MRSA clones in a city, country, and even among countries and continents has been identified by molecular techniques. We sought to determine whether clonal spread of MRSA has occurred in Trinidad and Tobago from all MRSA isolates collected between 2000 and 2001. METHODS: Clinical isolates of MRSA from three major hospitals in Trinidad and Tobago were identified by standard laboratory methods and analyzed using multiplex polymerase chain reaction (PCR) and pulsed-field gel electrophoresis (PFGE) after SmaI digestion. RESULTS: There was a 12.8% prevalence of MRSA in three major regional hospitals in Trinidad and Tobago. All 60 randomly selected MRSA strains from these hospitals produced similar PFGE banding patterns, suggesting a genetic relatedness among strains and that they belonged to a single clonal family. All isolates were negative for the Panton-Valentine leukocidin gene (pvl). These strains shared a PFGE banding pattern approximately (96%) the same as a Canadian strain called CMRSA-6 in the Canadian National Microbiology Laboratory database. CONCLUSIONS: We conclude that only one major PFGE genotype of MRSA clone is circulating among the three major regional hospitals in Trinidad and Tobago suggesting one of three possible scenarios of microevolution: (1) all were from the dissemination of a single epidemic MRSA clone prevailing in these hospitals in Trinidad and Tobago; or (2) MRSA in Trinidad and Tobago is evolving more slowly than in other countries; or (3) that if other MRSA clones have been present in Trinidad and Tobago, they have not persisted.     

北京分离株金黄色葡萄球菌基因型分析

目的对2000-2005年北京分离株金黄色葡萄球菌进行基因分型研究,了解其型别构成特征。方法分别对48株耐甲氧西林金黄色葡萄球菌（methicillin-resistant S．aureus,MRSA）和3株甲氧西林敏感金黄色葡萄球菌（methicillin-sensitive S．aureus,MSSA）进行spa基因序列分析及脉冲场凝胶电泳（pulsed field gel electrophoresis,PFGE）分析,并与多位点序列分析（multi locus sequencing typing,MLST）进行比较。结果在MRSA中,t030和t037为主要spa型别,分别占47．92％（23／48）和37．50％（18／48）;t002只在2005年菌株中出现,占14．58％（7／48）。3株MSSA型别分别为t377和t304。PFGE按带型差异及相似度分析,将所有菌株分为5组11型。结论t030和t037是MRSA临床分离株中的spa优势型别,A组和D组为PFGE优势型别。合理使用MLST、PFGE和spa分型方法对我国MRSA防治具有重要意义。     

The evolution of methicillin resistance in Staphylococcus aureus: similarity of genetic backgrounds in historically early methicillin-susceptible and -resistant isolates and contemporary epidemic clones

The key genetic component of methicillin resistance, the mecA determinant, is not native to Staphylococcus aureus. Thus, the evolution of methicillin-resistant S. aureus (MRSA) must have begun with the acquisition of the mecA determinant from an unknown heterologous source some time before the first reported appearance of MRSA isolates in clinical specimens in the U.K. and Denmark (in the early 1960s). We compared the genetic backgrounds and phenotypes of a group of methicillin-susceptible S. aureus (MSSA) isolates to the properties of MRSA strains isolated in Denmark and the U.K. during the same time period, and also to the genetic profiles of contemporary epidemic clones of MRSA. All early MRSA isolates resembled a large group of the early MSSA blood isolates in phenotypic and genetic properties, including phage group, antibiotype (resistance to penicillin, streptomycin, and tetracycline), pulsed-field gel electrophoresis pattern, and spaA type and multilocus sequence type, strongly suggesting that the early MSSA examined here represented the progeny of a strain that served as one of the first S. aureus recipients of the methicillin-resistance determinant in Europe. The genetic background of this group of early MSSA isolates was also very similar to that of the widely disseminated contemporary "Iberian clone" of MRSA, suggesting that genetic determinants present in early MSSA and essential for some aspects of the epidemicity and/or virulence of these strains may have been retained by this highly successful contemporary MRSA lineage.     

Evolution of erythromycin-resistant Streptococcus pneumoniae from Asian countries that contains erm(B) and mef(A) genes

To investigate the genetic characteristics of erythromycin-resistant Streptococcus pneumoniae in Asian countries, 110 pneumococcal isolates were analyzed by multilocus sequence typing (MLST). MLST analysis showed that 2 clonal complexes--CC236 (Taiwan19F-14 clone) and CC81 (Spain23F-1 clone)--are the major lineages of erythromycin-resistant S. pneumoniae in the Asian region. Pneumococcal isolates containing both the erm(B) and the mef(A) genes are thought to have originated from the Taiwan19F-14 clone containing the mef(A) gene, after introduction of the erm(B) gene. Further evolution of this variant clone has generated resistant strains with different sequence types. Dissemination of these variant clones of the Taiwan19F-14 could be the main reason for the high frequency of pneumococcal isolates containing both erm(B) and mef(A) in some Asian countries. Data suggest that the high prevalence of erythromycin-resistant S. pneumoniae in the Asian region is partly due to the clonal spread of a few multidrug-resistant clones.     

江苏地区金黄色葡萄球菌耐药及分子分型研究

目的 研究江苏省不同地区金葡菌的耐药性、耐药基因和分子分型的特征联系。方法 收集2017年全省各市哨点医院金黄色葡萄球菌227株,采用微量肉汤稀释法测定15种抗生素最小抑菌浓度, PCR方法检测7种不同的耐药基因,并用多位点序列分型(multilocus sequence typing MLST)进行分子分型。结果 本研究中金葡菌对青霉素、红霉素、头孢西丁、苯唑西林、克林霉素、左氧氟沙星、四环素、庆大霉素的耐药率较高,而对替考拉宁、利福平、复方新诺明、达托霉素、利奈唑胺以及呋喃妥因多呈敏感,耐药率均低于10%,且所有菌株均对万古霉素敏感。共计检出117株MRSA,MRSA平均耐药种数和平均耐药基因携带数高于MSSA(P< 0.05)。江苏省苏北地区的菌株耐药种数要高于苏中和苏南地区(P< 0.05),而苏中地区的耐药基因平均携带数高于苏南地区(P< 0.05)。在227株金葡菌中发现了60个ST型,以ST59和ST398分布数量最多,并有29个新ST型。成簇菌株中,CC59的平均耐药种数和MRSA检出率均高于其他克隆群。结论 江苏省内不同地区的金葡菌耐药率和耐药机制存在差异,且以CC59克隆群耐药情况最为严峻。     

Clonal types and antimicrobial resistance profiles of methicillin-resistant Staphylococcus aureus isolates from hospitals in south Brazil

In the present study were evaluated the DNA macrorestriction profile and SCCmec types for nine multi-resistant MRSA selected. Also antimicrobial susceptibility testing by disk diffusion method was evaluated for 68 MRSA isolates against 12 antimicrobial agents. The isolates were recovered from blood culture collected from hospitalized patients in three hospitals of Porto Alegre, Brazil. PFGE and PCR for mecA and SCCmec I, II, III, IV types genes were done on selected nine isolates with susceptibility only to vancomycin, teicoplanin and linezolid. Two clone profiles, with five subtypes, were demonstrated among multi-resistant MRSA analyzed. Eight isolates showed harbor SCCmec type III and one isolate was not typeable. The knowledge of SCCmec type, clone and antimicrobial profiles among S. aureus is essential mainly to prevention and control of dissemination of the antimicrobial resistance.