社区获得性甲氧西林耐药金黄色葡萄球菌的分子特征及毒力因子研究
|
摘要
[摘要]目的：了解2008年上海市儿童医学中心、上海市儿童医院、复旦大学附属儿科医院3家医院甲氧西林耐药金黄色葡萄球菌(MRSA)临床分离株中的PV杀白细胞素(PVL)基因分布、葡萄球菌mec盒式染色体(SCCmec)分型、多位点基因序列类型(MLST)分型及药物敏感性,并与同期甲氧西林敏感金黄色葡萄球菌(MSSA)的PVL基因分布及药物敏感性进行比较,分析社区获得性MRSA (CA-MRSA)的分子特征、毒力因子和药物敏感性。方法：对上海市三家儿童医院98株MRSA和49株MSSA临床分离株,用聚合酶链反应(PCR)方法筛选PVL基因阳性菌株并进行序列测定；用多重PCR方法对所有MRSA菌株进行SCCmec分型；用PCR方法通过金黄色葡萄球菌(SA)的7对看家基因(arcC、aroE、glpF、gmk、pta、tpi、yqi)对98株MRSA进行测序,通过MLST网站(http://www.mlst.net/)进行序列比对得出序列类型(STs)型别,eBURST软件分析各型在菌株克隆复合体(CC)内的相关性；用琼脂稀释法测定MRSA和MSSA对14种常用抗菌药物的敏感性(MIC)。结果：三家儿童医院的98株MRSA临床分离株中有6株PVL基因阳性,阳性率为6.1%(6／98),其中上海市儿童医学中心1株,上海市儿童医院4株,复旦大学附属儿科医院1株。49株MSSA临床分离株中有2株PVL基因阳性,阳性率为4.1%(2／49),该2株均分离自上海市儿童医院。所有MRSA临床分离株中发现4个SCCmec型别,其中Ⅱ型4株,占4.1%(4／98)；Ⅲ型23株,占23.5%(23／98)；Ⅳ型52株,占53%(52／98)；Ⅴ型15株,占15.3%(15／98)；4株(4.1%)未能分型。上海市儿童医学中心和上海市儿童医院以SCCmecⅣ型、Ⅴ型为主,复旦大学附属儿科医院则以SCCmecⅢ型多见。PVL基因阳性的6株MRSA中,有1株为SCCmecⅣ型,来自上海市儿童医院,余5株为SCCmecⅤ型。98株MRSA临床分离株中发现11个ST型别,其中ST59型33株,占33.7%(33／98)；ST239型29株,占29.6%(29／98)；ST5型12株,占12.2%(12／98)；ST88型9株,占9.2%(9／98)；STl型8株,占8.2%(8／98)；ST8型2株,占2%(2／98)；ST7、ST9、ST22、ST45、ST910各有一株,均占1%(1／98)。上海市儿童医学中心和上海市儿童医院多见ST59型,复旦大学附属儿科医院以ST239型为主。此外上海市儿童医院菌株中暂未发现ST239型存在。PVL基因阳性的6株MRSA中,3株为ST88-SCCmecⅤ,2株为ST59-SCCmecⅤ,1株为ST59-SCCmecⅣ。SCCmec分型中,4株Ⅱ型为ST5；23株Ⅲ型为ST239；52株Ⅳ型中30株为ST59,ST5和ST1各8株,ST8和ST88各2株,ST22和ST910各1株；15株Ⅴ型中6株为ST239、5株ST88、3株ST59、1株ST45。eBURST软件分析结果显示,ST8、ST239属于ST8 CC,ST1属于ST15 CC,ST910属于ST 30 CC,ST59、ST5、ST88、ST45、ST22、ST9、ST7均为各自CC的起源型别。MIC结果显示MSSA对药物的敏感性普遍高于MRSA,二者均对万古霉素、替考拉宁、利奈唑胺保持高度敏感,未发现万古霉素耐药株。PVL基因阳性株较阴性株对多种非β内酰胺类抗菌药更为敏感,但两者均对β内酰胺类、大环内酯类和克林霉素高度耐药,对庆大霉素、四环素的敏感性大致相仿。67株SCCmecⅣ、Ⅴ型MRSA对红霉素、克林霉素高度耐药,耐药率分别为92.5%和76.1%；对于其他非β内酰胺类抗菌药,67株SCCmecⅣ、V型MRSA的耐药率分别为：庆大霉素26.9%、利福平9%、四环素34.3%、左氧氟沙星14.9%、环丙沙星22.4%,均比27株SCCmecⅡ、Ⅲ型MRSA更为敏感；二者均对复方磺胺甲噁唑高度敏感。结论：在上海市三家儿童医院中,MRSA临床分离株的PVL基因阳性率相对较低,若仅以此作为CA-MRSA的标志可能漏检相当部分菌株。SCCmec分型可有效区分CA-MRSA和HA-MRSA。SCCmecⅣ型、V型菌株为CA-MRSA,可在院内传播,形成小范围内的流行,甚至暴发。SCCmecⅡ、Ⅲ型菌株的ST分型呈现高度克隆一致性,而SCCmecⅣ、Ⅴ型菌株中存在多种ST分型,并且分子量越小的SCCmec,相关的ST分型类别越多。CA-MRSA对多种非β内酰胺类抗菌药的敏感性显著高于HA-MRSA,多重耐药菌株的发生率较低,未发现万古霉素耐药株。
Objective The present study is aimed to investigate the distribution of Panton-Valentine leucocidin(PVL) gene, staphylococcal cassette chromosome mec (SCCmec) type, multilocus sequence types(MLST) type and the antimicrobial susceptibility of clinical isolates of Methicillin-resistant Staphylococcus aureus (MRSA) in three children's hospitals (Shanghai Children's Medical Center, Shanghai Children's Hospital, Children's Hospital of Fudan University) in 2008, and comparing the prevalence of PVL gene and antimicrobial susceptibility of methicillin-susceptible Staphylococcus aureus(MSSA)in the same periods, to analyze the molecular characteristic, the virulence factors and antimicrobial resistance of Community-acquired MRSA(CA-MRSA).
Methods ninety eight non-duplicate strains of MRSA and forty nine non-duplicate stains of MSSA isolated from the three Children's hospitals in Shanghai in 2008 were investigated. PVL genes were detected by polymerase chain reaction (PCR). The genotypes of SCCmec of the MRSA isolates were confirmed by multiplex PCR. The allelic profile of each strain was obtained by sequencing internal fragments of seven housekeeping genes (arcC, aroE, glpF, gmk, pta, tpi, and yqiL) and entering them on the MLST home page (http://saureus.mlst.net), where seven numbers depicting the allelic profile were assigned which defined an ST, at last the algorithm eBURST was used to identify groups of clonal complex (CC) and to predict the ancestral genotype of each group and the most parsimonious patterns of descent from the corresponding ancestor. The minimal inhibitory concentrations (MICs) of fourteen antibiotics for all isolates were determined by agar dilution method.
Results Among ninety eight isolates of MRSA, the PVL genes were found in 6.1%(6/98) of the MRSA strains, in which one isolate was from Shanghai Children's Medical Center, four isolates from Shanghai Children's Hospital, one isolate from Children's Hospital of Fudan University. In contrast, the PVL genes were found in 4.1%(2/49) of the MSSA strains, two isolates carring PVL genes were from Shanghai Children's Hospital, Whereas all the other isolates were negative for PVL genes. Among ninety eight isolates of MRSA,4.1%(4/98),23.5%(23/98),53%(52/98) and 15.3%(15/98) of the strains harboured SCCmec typesⅡ,Ⅲ,ⅣandⅤ, respectively, the remaining four isolates (4.1%) presented a unique SCCmec pattern that could not be classified to any known types by the employed typing assays. The SCCmec typeⅣand SCCmec type V were predominant in Shanghai Children's Medical Center and Shanghai Children's Hospital, while the SCCmec typeⅢwas predominant in Children's Hospital of Fudan University. Among six PVL gene-positive isolates of MRSA, one isolate which was from Shanghai Children's Hospital harbored SCCmec typeⅣ, the others were SCCmec typeⅤ. Eleven sequence types (STs) were identified among ninety eight isolates. The ST59 and ST239 isolates predominated in three Children's hospital in Shanghai and accounted for 33.7%(33/98) and 29.6%(29/98) respectively, ST5, ST88 and ST1 were not uncommon and accounted for 12.2%(12/98),9.2%(9/98) and 8.2%(8/98) respectively, the rest of the STs, such as ST8, ST7, ST9, ST22, ST45, ST910 were found in 1%-2% of the strains.ST59 was prevalent in Shanghai Children's Medical Center and Shanghai Children's Hospital, while ST239 was dominated in Children's Hospital of Fudan University. In addition, ST239 was not found in Shanghai Children's Hospital. Among six PVL gene-positive isolates of MRSA, three, two and one belonged to ST88-SCCmecⅤ, ST59-SCCmecⅤand ST59-SCCmecⅣrespectively. Combining the ST and SCCmec type, fourteen clone were identified, the predominant clone was ST59-SCCmecⅣ(thirty strains) and ST239-SCCmecⅢ(twenty tree strains), followed by ST5-SCCmecⅣand STl-SCCmecⅣ(eight strains for each clone), ST239-SCCmecⅤ(six strains), ST88-SCCmecⅤ(five strains), ST5-SCCmecⅡ(four strains), ST59-SCCmecⅤ(three strains), ST8-SCCmecⅣand ST88-SCCmecⅣ(two strains for each clone), ST22-SCCmecⅣ, ST910-SCCmecⅣand ST45-SCCmecⅤ(one strain for each clone). The eBURST analyses distributed the MRSA isolates into several CCs. ST8 and ST239 belonged to ST8 CC, ST1 belonged to ST15 CC, ST910 belonged to ST 30 CC, ST59, ST5, ST88, ST45, ST22, ST9 and ST7 were the origin of their own CCs. The results of MIC showed that the MSSA isolates were more susceptible than the MRSA isolates to fourteen antimicrobial agents tested, All strains including MSSA and MRSA were susceptible to vancomycin, teicoplanin and linezolid, no vancomycin-resistant strain was found. PVL gene-positive strains were much more susceptible to non-β-lactam antibiotics than those without PVL genes, both of them were highly resistant toβ-lactam antibiotics, macrolides and clindamycin, they had fairly susceptibility to gentamicin and tetracycline. Among sixty seven strains of MRSA harboring SCCmec type IV or SCCmec type V, the resistant rates to erythromycin and clindamycin were high, with 92.5% and 76.1% respectively. The resistant rates to other non-β-lactam antibiotics of sixty seven strains of MRSA were as follows: gentamicin 26.9%, rifampicin 9%, tetracycline 34.3%, levofloxacin 14.9%, ciprofloxacin 22.4%, which were more susceptible than twenty seven strains of MRSA harboring SCCmec typeⅡor SCCmec typeⅢ, both of them remained highly sensitivety to SMZ-TMP.
Conclusions Among three children's hospitals in Shanghai, the PVL gene-positive rate of MRSA isolates was relatively low, however,a considerable part of CA-MRSA strains might be undetected if PVL gene was treated as a sign of CA-MRSA. The genotypes of SCCmec could effectively distinguish CA-MRSA from HA-MRSA. SCCmec typeⅣand SCCmec typeⅤbelonged to CA-MRSA, which could spread among hospitals to form a small scale epidemic, even outbreaks. The STs of SCCmec type Hand SCCmec typeⅢstrains showed a high degree of consistency, while the SCCmec typeⅣand SCCmec type V strains had a variety of STs, furthermore, the smaller the SCCmec the more STs were associated with it. CA-MRSA was susceptible to various non-β-lactam antibiotics, with the low incidence of multidrug resistance. No vancomycin-resistant strain was found.
Objective The present study is aimed to investigate the distribution of Panton-Valentine leucocidin(PVL) gene, staphylococcal cassette chromosome mec (SCCmec) type, multilocus sequence types(MLST) type and the antimicrobial susceptibility of clinical isolates of Methicillin-resistant Staphylococcus aureus (MRSA) in three children's hospitals (Shanghai Children's Medical Center, Shanghai Children's Hospital, Children's Hospital of Fudan University) in 2008, and comparing the prevalence of PVL gene and antimicrobial susceptibility of methicillin-susceptible Staphylococcus aureus(MSSA)in the same periods, to analyze the molecular characteristic, the virulence factors and antimicrobial resistance of Community-acquired MRSA(CA-MRSA).
Methods ninety eight non-duplicate strains of MRSA and forty nine non-duplicate stains of MSSA isolated from the three Children's hospitals in Shanghai in 2008 were investigated. PVL genes were detected by polymerase chain reaction (PCR). The genotypes of SCCmec of the MRSA isolates were confirmed by multiplex PCR. The allelic profile of each strain was obtained by sequencing internal fragments of seven housekeeping genes (arcC, aroE, glpF, gmk, pta, tpi, and yqiL) and entering them on the MLST home page (http://saureus.mlst.net), where seven numbers depicting the allelic profile were assigned which defined an ST, at last the algorithm eBURST was used to identify groups of clonal complex (CC) and to predict the ancestral genotype of each group and the most parsimonious patterns of descent from the corresponding ancestor. The minimal inhibitory concentrations (MICs) of fourteen antibiotics for all isolates were determined by agar dilution method.
Results Among ninety eight isolates of MRSA, the PVL genes were found in 6.1%(6/98) of the MRSA strains, in which one isolate was from Shanghai Children's Medical Center, four isolates from Shanghai Children's Hospital, one isolate from Children's Hospital of Fudan University. In contrast, the PVL genes were found in 4.1%(2/49) of the MSSA strains, two isolates carring PVL genes were from Shanghai Children's Hospital, Whereas all the other isolates were negative for PVL genes. Among ninety eight isolates of MRSA,4.1%(4/98),23.5%(23/98),53%(52/98) and 15.3%(15/98) of the strains harboured SCCmec typesⅡ,Ⅲ,ⅣandⅤ, respectively, the remaining four isolates (4.1%) presented a unique SCCmec pattern that could not be classified to any known types by the employed typing assays. The SCCmec typeⅣand SCCmec type V were predominant in Shanghai Children's Medical Center and Shanghai Children's Hospital, while the SCCmec typeⅢwas predominant in Children's Hospital of Fudan University. Among six PVL gene-positive isolates of MRSA, one isolate which was from Shanghai Children's Hospital harbored SCCmec typeⅣ, the others were SCCmec typeⅤ. Eleven sequence types (STs) were identified among ninety eight isolates. The ST59 and ST239 isolates predominated in three Children's hospital in Shanghai and accounted for 33.7%(33/98) and 29.6%(29/98) respectively, ST5, ST88 and ST1 were not uncommon and accounted for 12.2%(12/98),9.2%(9/98) and 8.2%(8/98) respectively, the rest of the STs, such as ST8, ST7, ST9, ST22, ST45, ST910 were found in 1%-2% of the strains.ST59 was prevalent in Shanghai Children's Medical Center and Shanghai Children's Hospital, while ST239 was dominated in Children's Hospital of Fudan University. In addition, ST239 was not found in Shanghai Children's Hospital. Among six PVL gene-positive isolates of MRSA, three, two and one belonged to ST88-SCCmecⅤ, ST59-SCCmecⅤand ST59-SCCmecⅣrespectively. Combining the ST and SCCmec type, fourteen clone were identified, the predominant clone was ST59-SCCmecⅣ(thirty strains) and ST239-SCCmecⅢ(twenty tree strains), followed by ST5-SCCmecⅣand STl-SCCmecⅣ(eight strains for each clone), ST239-SCCmecⅤ(six strains), ST88-SCCmecⅤ(five strains), ST5-SCCmecⅡ(four strains), ST59-SCCmecⅤ(three strains), ST8-SCCmecⅣand ST88-SCCmecⅣ(two strains for each clone), ST22-SCCmecⅣ, ST910-SCCmecⅣand ST45-SCCmecⅤ(one strain for each clone). The eBURST analyses distributed the MRSA isolates into several CCs. ST8 and ST239 belonged to ST8 CC, ST1 belonged to ST15 CC, ST910 belonged to ST 30 CC, ST59, ST5, ST88, ST45, ST22, ST9 and ST7 were the origin of their own CCs. The results of MIC showed that the MSSA isolates were more susceptible than the MRSA isolates to fourteen antimicrobial agents tested, All strains including MSSA and MRSA were susceptible to vancomycin, teicoplanin and linezolid, no vancomycin-resistant strain was found. PVL gene-positive strains were much more susceptible to non-β-lactam antibiotics than those without PVL genes, both of them were highly resistant toβ-lactam antibiotics, macrolides and clindamycin, they had fairly susceptibility to gentamicin and tetracycline. Among sixty seven strains of MRSA harboring SCCmec type IV or SCCmec type V, the resistant rates to erythromycin and clindamycin were high, with 92.5% and 76.1% respectively. The resistant rates to other non-β-lactam antibiotics of sixty seven strains of MRSA were as follows: gentamicin 26.9%, rifampicin 9%, tetracycline 34.3%, levofloxacin 14.9%, ciprofloxacin 22.4%, which were more susceptible than twenty seven strains of MRSA harboring SCCmec typeⅡor SCCmec typeⅢ, both of them remained highly sensitivety to SMZ-TMP.
Conclusions Among three children's hospitals in Shanghai, the PVL gene-positive rate of MRSA isolates was relatively low, however,a considerable part of CA-MRSA strains might be undetected if PVL gene was treated as a sign of CA-MRSA. The genotypes of SCCmec could effectively distinguish CA-MRSA from HA-MRSA. SCCmec typeⅣand SCCmec typeⅤbelonged to CA-MRSA, which could spread among hospitals to form a small scale epidemic, even outbreaks. The STs of SCCmec type Hand SCCmec typeⅢstrains showed a high degree of consistency, while the SCCmec typeⅣand SCCmec type V strains had a variety of STs, furthermore, the smaller the SCCmec the more STs were associated with it. CA-MRSA was susceptible to various non-β-lactam antibiotics, with the low incidence of multidrug resistance. No vancomycin-resistant strain was found.
引文
[1]Almer LS, Shortridge VD, Nilius AM, etal. Antimicrobial susceptibility and molecular characterization of community-acquired methicillin-resistant Staphylococcus aureus[J]. Diagnostic Microbiology and Infection Disease,2002,43:225-232.
[2]Purcell K, Fergie J. Epidemic of community-acquired methicillin-resistant Staphylococcus aureus infections:a 14-year study at Driscoll Children's Hospital [J]. Arch Pediatr Adolesc Med, 2005,159(10):980-985.
[3]Otter JA, French GL. Nosocomial transmission of community-associated methicillin-resistant Staphylococcus aureus:an emerging threat [J]. Lancet Infect Dis,2006,6(12):753-755.
[4]Vandenesch F, Naimi T, Enright MC, etal. Community-acquired methicillin-resistant Staphylococcus aureus carrying Panton-Valentine leukocidin genes:worldwide emergence[J]. Emerg Infect Dis,2003,9(8):978-984.
[5]Boyle-Vavra S, Daum RS. Community-acquired methicillin-resistant Staphylococcus aureus:the role of Panton-Valentine leukocidin [J]. Lab Invest,2007,87:3-9.
[6]McClure JA, Conly JM, Lau V, etal. Novel Multiplex PCR Assay for Detection of the Staphylococcal Virulence Marker Panton-Valentine Leukocidin Genes and Simultaneous Discrimination of Methicillin-Susceptible from-Resistant Staphylococci [J]. J Clin Microbiol,2006,44(3):1141-1144.
[7]余方友,陈增强,林晓梅,等.多重PCR对金黄色葡萄球菌杀白细胞素基因的检测[J].中华微生物学和免疫学杂志,2006,26(12)：1112-1115.
[8]叶慧.耐甲氧西林的金黄色葡萄球菌SCCmec分型及PVL基因的表达[D].四川：四川大学,2007：
[9]Nimmo GR, Coombs GW, Pearson PC, Methicillin-resistant Staphylococcus aureus in the Australian community:An evolving epidemic[J]. Med J Aust,2006,184(8):384-388.
[10]陆敏,张泓,乔蓉,等.儿童社区获得性耐甲氧西林金黄色葡萄球菌感染研究[J].临床儿科杂志,2009,27(6)：528-533.
[11]王辉,刘昱东,杜娜,等.2005年我国多中心苯唑西林耐药的金黄色葡萄球菌分子流行病学研究[J].中华检验医学杂志,2007,30(12)：1354-1359.
[12]周晓兰,李光辉,徐晓刚,等.杀白细胞毒素基因阳性MRSA的临床和实验研究[J].中国感染与化疗杂志,2008,8(3)：167-171.
[13]王磊.社区获得性耐甲氧西林金黄色葡萄球菌临床株遗传学分析[D].天津：天津医科大学,2008：
[14]Michael M. Idcp snapshots[J]. Infect Dis Clin Prac,2002,11(6):327.
[15]Piriyaporn C, Ito T, Ma X, et al. Staphylococcal cassette chromosome mec (SCCmec) typing of methicillin-resistant Staphylococcus aureus strains isolated in 11 Asian countries:a proposal for a new nomenclature for SCCmec elements[J]. Antimicrob Agents Chemother,2006,50(3):1001.
[16]Berglund C, Ito T, Ma XX, etal. Genetic diversity of methicillin-resistant staphylococcus aureus carring type Ⅳ SCCmec in Orebro country and the western region of Sweden[J]. Journal of Antimicrobial Chemotherapy,2009,63:32-41.
[17]International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC). Classification of Staphylococcal Cassette Chromosome mec (SCCmec):Guidelines for Reporting Novel SCCmec Elements[J]. Antimicrobial agents and chemotherapy,2009,53(12):4961-4967.
[18]Jemili-Ben MJ, Boutiba-Ben IB, Ben SR. Identification of staphylococcal cassette chromosome mec encoding methicillin resistance in Staphylococcus aureus isolates at Charles Nicolle Hospital of Tunis[J], Pathologie Biologie,2006,54:453-455.
[19]Maree CL, Daum RS, Boyle-Vavra S, etal. Community-associated methicillin-resistant Staphylococcus aureus isolates causing healthcare-associated infections[J]. Emerg Infect Dis,2007, 13(2):236-242.
[20]Gordon RJ, Lowy FD. Pathogenesis of Methicillin-Resistant Staphylococcus aureus Infection[J]. Clin Infect Dis,2008,46(suppl 5):350-359
[21]Oliveira DC, de Lencastre H.Multiplex PCR Strategy for Rapid Identification of Structural Types and Variants of the mec Element in Methicillin-Resistant Staphylococcus aureus[J]. Antimicrob Agents Chemother,2002,46(7):2155-2161.
[22]Zhang KY, McClure JA, Elsayed S.etal.Novel Multiplex PCR Assay for Characterization and Concomitant Subtyping of Staphylococcal Cassette Chromosome mec Types Ⅰ to Ⅴ in Methicillin-Resistant Staphylococcus aureus. Journal of Clinical Microbiology,2005, 43(10):5026-5033.
[23]Valsesia G, Rossi M, Bertschy S, etal. Emergence of SCCmec Type Ⅳ and SCCmec Type Ⅴ Methicillin-Resistant Staphylococcus aureus Containing the Panton-Valentine Leukocidin Genes in a Large Academic Teaching Hospital in Central Switzerland:External Invaders or Persisting Circulators?[J]. J Clin Microbiol,2010,48:720-727.
[24]Frindkin SK, Hageman JC, Morrison M, etal. Methicillin-resistant Staphylococcus aureus disease in three communities[J].N Engl J Med,2005,352(14):1436-1444.
[25]张文双.耐甲氧西林金黄色葡萄球菌的流行状况及其起源进化[J].中国当代儿科杂志,2008,10(3)：426-430.
[26]David MZ,Crawford SE, Boyle-Vavra S,etal. Contrasting pediatric and adult methicillin-resistant Staphylococcus aureus isolates [J]. Emerg Infect Dis,2006,12 (4):631-637.
[27]Takano T, Higuchi W, Otsuka T,etal. novel characteristics of community-acquired methicillin-resistant staphylococcus aureus strains belonging to multilocus sequence type 59 in Taiwan[J], Antimicrobial agents and chemotherapy,2008,52(3):837-845.
[28]Enright M C, Robinson D A, Randle G, et al. The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA) [J]. Proc Natl Acad Sci USA,2002,99(11):7687.
[29]Takano T, Higuchi W, Otsuka T, etal. novel characteristics of community-acquired methicillin-resistant staphylococcus aureus strains belonging to multilocus sequence type 59 in Taiwan[J], Antimicrobial agents and chemotherapy,2008,52(3):837-845.
[30]http://www.mlst.net/
[31]Denis O, Deplano A, De Beenhouwer H. Polyclonal emergence and importation of community—acquired methicillin—resistant Staphylococcus aureus strains harboring Panton—Valentine leucocidin genes in Belgium. J Antimicrob Chemother,2005,56: 1103-1106.
[32]Ma XX, Ito T, Chongtrakool P, etal. Predominance of clones carring Paton-Valentine leukocidin genes among methicillin-resistant Staphylococcus aureus strains isolated in Japanese hospitals from 1979 to 1985[J],J Clin Microbiol,2006,44(12):4515-4527.
[33]Deurenberg RH.Vin KC, kalenic S, etal. The molecular evolution of methicillin-resistant Staphylococcus aureus[J]. Clin Microbiol Infect,2007,13 (3):222-235.
[34]Geng WJ, Yang YH, Wu DJ, etal. Community-acquired, methicillin-resistant Staphylococcus aureus isolated from children with community-onset pneumonia in China[J]. Pediatr Pulmonol,2010,45(4):387-94.
[35]Robinson DA, Kearns AM, Holmes A, etal. Re-emergence of early pandemic Staphylococcus aurues as a community-acquired methicillin-resistant clone[J]. Lancet,2005,365(9466):1256-1258.
[36]闫笑梅,顾一心,何利华,等.耐甲氧西林金黄色葡萄球菌多位点序列分析[J],中华预防医学杂志,2009,43(2)：137-140.
[37]Wielders CL, Vriens MR, Brisse S, etal. In-vivo transfer of mecA DNA to Staphylococcus aureus[J]. Lancet,2001,357(9269):1674-1675.
[38]刘盈,张霞,马琳,等.北京地区儿童皮肤感染金黄色葡萄球菌的耐药研究[J].中华皮肤科杂志,2008,41(4)：214-216.
[39]马富艳,吴晓平,华春珍.儿童与成人感染金黄色葡萄球菌耐药性对比分析[J].中华医院感染学杂志,2007,17(03)：331-333.
[1]Klevens RM, Morrison MA, Nadle J, et al. Invasive methicillin-resistant Staphylococcus aureus Infections in the United States[J]. JAMA,2007,298:1763-1771.
[2]Karchmer AW, Bayer AS. Methicillin-Resistant Staphylococcus aureus:An Evolving Clinical Challenge[J]. Clin Infect Dis,2008,46(Suppl 5):342.
[3]Boucher HW, Corey GR. Epidemiology of Methicillin-Resistant Staphylococcus aureus[J]. Clin Infect Dis,2008,46(Suppl 5):344.
[4]Nathwani D, Morgan M, Masterton RG, et al. Guidelines for UK practice for the diagnosis and management of methicillin-resistant Staphylococcus aureus (MRSA) infections presenting in the community. J Antimicrob Chemother,2008; 61(5):976-994.
[5]Harbarth S, Francois P, Shrenzel J, et al. Community-associated Methicillin-resistant staphylococcus aureus, Switzerland. Emerging Infectious Diseases[J],2005,11(6):962-965.
[6]Wijaya L, Hsu LY, Kurup A. Community-associated Methicillin-resistant Staphulococcus aureus:Overview and Local Situation[R]. Ann Acad Med Singapore,2006,35:479-86.
[7]Miller LG, Diep BA. Colonization, Fomites, and Virulence:Rethinking the Pathogenesis of Community-Associated Methicillin-Resistant Staphylococcus aureus Infection [J]. Clin Inf Dis,2008,46(5):752-760.
[8]Hsu LY, Koh YL, Chlebicka NL, et al. Establishment of ST30 as the Predominant Clonal Yin-Ling Type among Community-Associated Methicillin-Resistant Staphylococcus aureus Isolates in Singapore[J]. J Clin Microbiol,2006,44,3:1090-1093.
[9]Stryjewski ME, Chamber HF. Skin and Soft-Tissue Infections Caused by Community-Acquired Methicillin-Resistant Staphylococcus aureus [J]. Clin Microbiol,2008,46(suppl 5):S368-S377.
[10]International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC). Classification of Staphylococcal Cassette Chromosome mec (SCCmec):Guidelines for Reporting Novel SCCmec Elements[J]. Antimicrobial agents and chemotherapy,2009,53(12):4961-4967
[11]Gordon RJ, Lowy FD. Pathogenesis of Methicillin-Resistant Staphylococcus aureus Infection[J]. Clin Infect Dis,2008,46(suppl 5):350-359.
[12]Memmi G, Filipe SR, Pinho MG, et al. Staphylococcus aureus PBP4 Is Essential for β-Lactam Resistance in Community-Acquired Methicillin-Resistant Strains[J]. Antimicrobial Agents And Chemotherapy,2008,52(11):3955-3966.
[13]Labandeira-Rey M, Couzon F, Boisset S, et al. Staphylococcus aureus Panton-Valentine leukocidin casuses necrotizing pneumonia [J]. Science,2007,315(5815):1130—1133.
[14]Voyich JM, Otto M, Mathema B, et al. Is Panton-Valentine leukocidin the major virulence determinant in community-associated methicillin-resistant Staphylococcus aureus disease?[J]. J Infect Dis,2006,194(12):1761-70.
[15]Otter JA, French GL. Nosocomial transmission of community-associated methicillin-resistant Staphylococcus aureus:an emerging threat [J]. Lancet Infect Dis,2006,6(12):753-755.
[16]Stryjewski ME, Chambers HF. Skin and Soft-Tissue Infections Caused by Community-Acquired Methicillin-Resistant Staphylococcus aureus[J]. Clin Infect Dis,2008,46(Suppl 5):368-377.
[17]Gillet Y, Issartel B, Vanhems P, et al. Association between Staphylococcus aureus strains carrying gene for Panton-Valentine leukocidin and highly lethal necrotising pneumonia in young immunocompetent patients[J],lancet,2002,359(9308):753-759.
[18]Rubinstein E, Kollef MH, Nathwani D. Pneumonia Caused by Methicillin-Resistant Staphylococcus aureus [J], Clin Infect Dis,2008,46(Suppl 5):378-385.
[19]Cosgrove SE, Fowler VG Jr.Management of Methicillin-Resistant Staphylococcus aureus Bacteremia [J], Clin Infect Dis,2008,46(Suppl 5):386-393.
[20]胡付品,吴湜,叶信予,等.头孢西丁纸片扩散法检测耐甲氧西林葡萄球菌[J].中国抗感染化疗杂志,2005,5(2)：86-88.
[21]Naimi T, LeDell KH, Como-Sabetti K, Borchardt SM, Boxrud DJ, Etienne J, et al. Community and healthcare-associated methicillin-resistant Staphylococcus aureus infections in Minnesota. JAMA. 2003:290:2976-84.
[22]Moellering RC Jr. Current Treatment Options for Community-Acquired Methicillin-Resistant Staphylococcus aureus Infection [J], Clin Infect Dis,2008,46:1032-1037.
[23]Dumitrescu 0, Boisset S, Badiou C, et al. Effect of antibiotics on Staphylococcus aureus producing Panton-Valentine leukocidin[J]. Antimicrob Agents Chemother,2007,51 (4):1515-15199.
[24]Dumitrescu O, Boisset S, Badiou C, et al. Effect of antibiotics on Staphylococcus aureus producing Panton-Valentine leukocidin[J]. Antimicrob Agents Chemother,2007,51(4):1515-15199.
[25]Sakoulas G, Moellering RC Jr. Increasing Antibiotic Resistance among Methicillin-Resistant Staphylococcus aureus Strains [J]. Clin Infect Dis,2008,46(Suppl 5):360-367.
[26]Fridkin SK. Vancomycin—intermediate and-resistant Staphylococcus aureus:What the infectious disease specialist needs to know. Clin Inf Dis,2001,32 (1):108—115.
[27]Cui LZ, Ma XX, Sato K, et al. Cell wall thickening is a common feature of vancomycin resistance in Staphylococcus aureus. J Clin Microbiol, 2003,41 (1):5—14.
[28]汪复.社区获得性甲氧西林耐药金黄色葡萄球菌[J].中国抗感染化疗杂 志,2005,5(6)：376-380.
[29]Dumitrescu O, Boisset S, Badiou C, et al. Effect of antibiotics on Staphylococcus aureus producing Panton-Valentine leukocidin[J]. Antimicrob Agents Chemother,2007,51(4):1515-15199.
[2]Purcell K, Fergie J. Epidemic of community-acquired methicillin-resistant Staphylococcus aureus infections:a 14-year study at Driscoll Children's Hospital [J]. Arch Pediatr Adolesc Med, 2005,159(10):980-985.
[3]Otter JA, French GL. Nosocomial transmission of community-associated methicillin-resistant Staphylococcus aureus:an emerging threat [J]. Lancet Infect Dis,2006,6(12):753-755.
[4]Vandenesch F, Naimi T, Enright MC, etal. Community-acquired methicillin-resistant Staphylococcus aureus carrying Panton-Valentine leukocidin genes:worldwide emergence[J]. Emerg Infect Dis,2003,9(8):978-984.
[5]Boyle-Vavra S, Daum RS. Community-acquired methicillin-resistant Staphylococcus aureus:the role of Panton-Valentine leukocidin [J]. Lab Invest,2007,87:3-9.
[6]McClure JA, Conly JM, Lau V, etal. Novel Multiplex PCR Assay for Detection of the Staphylococcal Virulence Marker Panton-Valentine Leukocidin Genes and Simultaneous Discrimination of Methicillin-Susceptible from-Resistant Staphylococci [J]. J Clin Microbiol,2006,44(3):1141-1144.
[7]余方友,陈增强,林晓梅,等.多重PCR对金黄色葡萄球菌杀白细胞素基因的检测[J].中华微生物学和免疫学杂志,2006,26(12)：1112-1115.
[8]叶慧.耐甲氧西林的金黄色葡萄球菌SCCmec分型及PVL基因的表达[D].四川：四川大学,2007：
[9]Nimmo GR, Coombs GW, Pearson PC, Methicillin-resistant Staphylococcus aureus in the Australian community:An evolving epidemic[J]. Med J Aust,2006,184(8):384-388.
[10]陆敏,张泓,乔蓉,等.儿童社区获得性耐甲氧西林金黄色葡萄球菌感染研究[J].临床儿科杂志,2009,27(6)：528-533.
[11]王辉,刘昱东,杜娜,等.2005年我国多中心苯唑西林耐药的金黄色葡萄球菌分子流行病学研究[J].中华检验医学杂志,2007,30(12)：1354-1359.
[12]周晓兰,李光辉,徐晓刚,等.杀白细胞毒素基因阳性MRSA的临床和实验研究[J].中国感染与化疗杂志,2008,8(3)：167-171.
[13]王磊.社区获得性耐甲氧西林金黄色葡萄球菌临床株遗传学分析[D].天津：天津医科大学,2008：
[14]Michael M. Idcp snapshots[J]. Infect Dis Clin Prac,2002,11(6):327.
[15]Piriyaporn C, Ito T, Ma X, et al. Staphylococcal cassette chromosome mec (SCCmec) typing of methicillin-resistant Staphylococcus aureus strains isolated in 11 Asian countries:a proposal for a new nomenclature for SCCmec elements[J]. Antimicrob Agents Chemother,2006,50(3):1001.
[16]Berglund C, Ito T, Ma XX, etal. Genetic diversity of methicillin-resistant staphylococcus aureus carring type Ⅳ SCCmec in Orebro country and the western region of Sweden[J]. Journal of Antimicrobial Chemotherapy,2009,63:32-41.
[17]International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC). Classification of Staphylococcal Cassette Chromosome mec (SCCmec):Guidelines for Reporting Novel SCCmec Elements[J]. Antimicrobial agents and chemotherapy,2009,53(12):4961-4967.
[18]Jemili-Ben MJ, Boutiba-Ben IB, Ben SR. Identification of staphylococcal cassette chromosome mec encoding methicillin resistance in Staphylococcus aureus isolates at Charles Nicolle Hospital of Tunis[J], Pathologie Biologie,2006,54:453-455.
[19]Maree CL, Daum RS, Boyle-Vavra S, etal. Community-associated methicillin-resistant Staphylococcus aureus isolates causing healthcare-associated infections[J]. Emerg Infect Dis,2007, 13(2):236-242.
[20]Gordon RJ, Lowy FD. Pathogenesis of Methicillin-Resistant Staphylococcus aureus Infection[J]. Clin Infect Dis,2008,46(suppl 5):350-359
[21]Oliveira DC, de Lencastre H.Multiplex PCR Strategy for Rapid Identification of Structural Types and Variants of the mec Element in Methicillin-Resistant Staphylococcus aureus[J]. Antimicrob Agents Chemother,2002,46(7):2155-2161.
[22]Zhang KY, McClure JA, Elsayed S.etal.Novel Multiplex PCR Assay for Characterization and Concomitant Subtyping of Staphylococcal Cassette Chromosome mec Types Ⅰ to Ⅴ in Methicillin-Resistant Staphylococcus aureus. Journal of Clinical Microbiology,2005, 43(10):5026-5033.
[23]Valsesia G, Rossi M, Bertschy S, etal. Emergence of SCCmec Type Ⅳ and SCCmec Type Ⅴ Methicillin-Resistant Staphylococcus aureus Containing the Panton-Valentine Leukocidin Genes in a Large Academic Teaching Hospital in Central Switzerland:External Invaders or Persisting Circulators?[J]. J Clin Microbiol,2010,48:720-727.
[24]Frindkin SK, Hageman JC, Morrison M, etal. Methicillin-resistant Staphylococcus aureus disease in three communities[J].N Engl J Med,2005,352(14):1436-1444.
[25]张文双.耐甲氧西林金黄色葡萄球菌的流行状况及其起源进化[J].中国当代儿科杂志,2008,10(3)：426-430.
[26]David MZ,Crawford SE, Boyle-Vavra S,etal. Contrasting pediatric and adult methicillin-resistant Staphylococcus aureus isolates [J]. Emerg Infect Dis,2006,12 (4):631-637.
[27]Takano T, Higuchi W, Otsuka T,etal. novel characteristics of community-acquired methicillin-resistant staphylococcus aureus strains belonging to multilocus sequence type 59 in Taiwan[J], Antimicrobial agents and chemotherapy,2008,52(3):837-845.
[28]Enright M C, Robinson D A, Randle G, et al. The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA) [J]. Proc Natl Acad Sci USA,2002,99(11):7687.
[29]Takano T, Higuchi W, Otsuka T, etal. novel characteristics of community-acquired methicillin-resistant staphylococcus aureus strains belonging to multilocus sequence type 59 in Taiwan[J], Antimicrobial agents and chemotherapy,2008,52(3):837-845.
[30]http://www.mlst.net/
[31]Denis O, Deplano A, De Beenhouwer H. Polyclonal emergence and importation of community—acquired methicillin—resistant Staphylococcus aureus strains harboring Panton—Valentine leucocidin genes in Belgium. J Antimicrob Chemother,2005,56: 1103-1106.
[32]Ma XX, Ito T, Chongtrakool P, etal. Predominance of clones carring Paton-Valentine leukocidin genes among methicillin-resistant Staphylococcus aureus strains isolated in Japanese hospitals from 1979 to 1985[J],J Clin Microbiol,2006,44(12):4515-4527.
[33]Deurenberg RH.Vin KC, kalenic S, etal. The molecular evolution of methicillin-resistant Staphylococcus aureus[J]. Clin Microbiol Infect,2007,13 (3):222-235.
[34]Geng WJ, Yang YH, Wu DJ, etal. Community-acquired, methicillin-resistant Staphylococcus aureus isolated from children with community-onset pneumonia in China[J]. Pediatr Pulmonol,2010,45(4):387-94.
[35]Robinson DA, Kearns AM, Holmes A, etal. Re-emergence of early pandemic Staphylococcus aurues as a community-acquired methicillin-resistant clone[J]. Lancet,2005,365(9466):1256-1258.
[36]闫笑梅,顾一心,何利华,等.耐甲氧西林金黄色葡萄球菌多位点序列分析[J],中华预防医学杂志,2009,43(2)：137-140.
[37]Wielders CL, Vriens MR, Brisse S, etal. In-vivo transfer of mecA DNA to Staphylococcus aureus[J]. Lancet,2001,357(9269):1674-1675.
[38]刘盈,张霞,马琳,等.北京地区儿童皮肤感染金黄色葡萄球菌的耐药研究[J].中华皮肤科杂志,2008,41(4)：214-216.
[39]马富艳,吴晓平,华春珍.儿童与成人感染金黄色葡萄球菌耐药性对比分析[J].中华医院感染学杂志,2007,17(03)：331-333.
[1]Klevens RM, Morrison MA, Nadle J, et al. Invasive methicillin-resistant Staphylococcus aureus Infections in the United States[J]. JAMA,2007,298:1763-1771.
[2]Karchmer AW, Bayer AS. Methicillin-Resistant Staphylococcus aureus:An Evolving Clinical Challenge[J]. Clin Infect Dis,2008,46(Suppl 5):342.
[3]Boucher HW, Corey GR. Epidemiology of Methicillin-Resistant Staphylococcus aureus[J]. Clin Infect Dis,2008,46(Suppl 5):344.
[4]Nathwani D, Morgan M, Masterton RG, et al. Guidelines for UK practice for the diagnosis and management of methicillin-resistant Staphylococcus aureus (MRSA) infections presenting in the community. J Antimicrob Chemother,2008; 61(5):976-994.
[5]Harbarth S, Francois P, Shrenzel J, et al. Community-associated Methicillin-resistant staphylococcus aureus, Switzerland. Emerging Infectious Diseases[J],2005,11(6):962-965.
[6]Wijaya L, Hsu LY, Kurup A. Community-associated Methicillin-resistant Staphulococcus aureus:Overview and Local Situation[R]. Ann Acad Med Singapore,2006,35:479-86.
[7]Miller LG, Diep BA. Colonization, Fomites, and Virulence:Rethinking the Pathogenesis of Community-Associated Methicillin-Resistant Staphylococcus aureus Infection [J]. Clin Inf Dis,2008,46(5):752-760.
[8]Hsu LY, Koh YL, Chlebicka NL, et al. Establishment of ST30 as the Predominant Clonal Yin-Ling Type among Community-Associated Methicillin-Resistant Staphylococcus aureus Isolates in Singapore[J]. J Clin Microbiol,2006,44,3:1090-1093.
[9]Stryjewski ME, Chamber HF. Skin and Soft-Tissue Infections Caused by Community-Acquired Methicillin-Resistant Staphylococcus aureus [J]. Clin Microbiol,2008,46(suppl 5):S368-S377.
[10]International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC). Classification of Staphylococcal Cassette Chromosome mec (SCCmec):Guidelines for Reporting Novel SCCmec Elements[J]. Antimicrobial agents and chemotherapy,2009,53(12):4961-4967
[11]Gordon RJ, Lowy FD. Pathogenesis of Methicillin-Resistant Staphylococcus aureus Infection[J]. Clin Infect Dis,2008,46(suppl 5):350-359.
[12]Memmi G, Filipe SR, Pinho MG, et al. Staphylococcus aureus PBP4 Is Essential for β-Lactam Resistance in Community-Acquired Methicillin-Resistant Strains[J]. Antimicrobial Agents And Chemotherapy,2008,52(11):3955-3966.
[13]Labandeira-Rey M, Couzon F, Boisset S, et al. Staphylococcus aureus Panton-Valentine leukocidin casuses necrotizing pneumonia [J]. Science,2007,315(5815):1130—1133.
[14]Voyich JM, Otto M, Mathema B, et al. Is Panton-Valentine leukocidin the major virulence determinant in community-associated methicillin-resistant Staphylococcus aureus disease?[J]. J Infect Dis,2006,194(12):1761-70.
[15]Otter JA, French GL. Nosocomial transmission of community-associated methicillin-resistant Staphylococcus aureus:an emerging threat [J]. Lancet Infect Dis,2006,6(12):753-755.
[16]Stryjewski ME, Chambers HF. Skin and Soft-Tissue Infections Caused by Community-Acquired Methicillin-Resistant Staphylococcus aureus[J]. Clin Infect Dis,2008,46(Suppl 5):368-377.
[17]Gillet Y, Issartel B, Vanhems P, et al. Association between Staphylococcus aureus strains carrying gene for Panton-Valentine leukocidin and highly lethal necrotising pneumonia in young immunocompetent patients[J],lancet,2002,359(9308):753-759.
[18]Rubinstein E, Kollef MH, Nathwani D. Pneumonia Caused by Methicillin-Resistant Staphylococcus aureus [J], Clin Infect Dis,2008,46(Suppl 5):378-385.
[19]Cosgrove SE, Fowler VG Jr.Management of Methicillin-Resistant Staphylococcus aureus Bacteremia [J], Clin Infect Dis,2008,46(Suppl 5):386-393.
[20]胡付品,吴湜,叶信予,等.头孢西丁纸片扩散法检测耐甲氧西林葡萄球菌[J].中国抗感染化疗杂志,2005,5(2)：86-88.
[21]Naimi T, LeDell KH, Como-Sabetti K, Borchardt SM, Boxrud DJ, Etienne J, et al. Community and healthcare-associated methicillin-resistant Staphylococcus aureus infections in Minnesota. JAMA. 2003:290:2976-84.
[22]Moellering RC Jr. Current Treatment Options for Community-Acquired Methicillin-Resistant Staphylococcus aureus Infection [J], Clin Infect Dis,2008,46:1032-1037.
[23]Dumitrescu 0, Boisset S, Badiou C, et al. Effect of antibiotics on Staphylococcus aureus producing Panton-Valentine leukocidin[J]. Antimicrob Agents Chemother,2007,51 (4):1515-15199.
[24]Dumitrescu O, Boisset S, Badiou C, et al. Effect of antibiotics on Staphylococcus aureus producing Panton-Valentine leukocidin[J]. Antimicrob Agents Chemother,2007,51(4):1515-15199.
[25]Sakoulas G, Moellering RC Jr. Increasing Antibiotic Resistance among Methicillin-Resistant Staphylococcus aureus Strains [J]. Clin Infect Dis,2008,46(Suppl 5):360-367.
[26]Fridkin SK. Vancomycin—intermediate and-resistant Staphylococcus aureus:What the infectious disease specialist needs to know. Clin Inf Dis,2001,32 (1):108—115.
[27]Cui LZ, Ma XX, Sato K, et al. Cell wall thickening is a common feature of vancomycin resistance in Staphylococcus aureus. J Clin Microbiol, 2003,41 (1):5—14.
[28]汪复.社区获得性甲氧西林耐药金黄色葡萄球菌[J].中国抗感染化疗杂 志,2005,5(6)：376-380.
[29]Dumitrescu O, Boisset S, Badiou C, et al. Effect of antibiotics on Staphylococcus aureus producing Panton-Valentine leukocidin[J]. Antimicrob Agents Chemother,2007,51(4):1515-15199.