血流感染金黄色葡萄球菌分子特征及毒力基因分析
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摘要
目的了解新疆军区总医院血流感染金黄色葡萄球菌临床分离株的分子特征,检测毒力基因携带情况及其对临床常用抗菌药物的耐药性。方法收集2012-2016年新疆军区总医院不同血流感染患者分离的53株金黄色葡萄球菌,应用聚合酶链式反应(Polymerase Chain Reaction,PCR)鉴定甲氧西林耐药性;应用SCCmec分型、spa分型、多位点序列分型(multilocus sequence typing,MLST)等分子分型方法对金黄色葡萄球菌进行分析,PCR检测pvl等21种毒力基因,分析不同克隆毒力基因携带情况,观察菌株对临床常用抗菌药物的耐药性。结果 53株金黄色葡萄球菌中30株为耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA),23株为甲氧西林敏感金黄色葡萄球菌(methicillin-susceptible Staphylococcus aureus,MSSA)。30株MRSA菌分为4种ST型和4种spa型,3种SCCmec型,优势流行克隆为MRSA-ST239-t030-SCCmecⅢ占90.0%(27/30)同时鉴定出3株社区获得性MRSA。23株MSSA鉴定出10种ST型、14种spa型,主要流行克隆为ST22-t309,检出4株动物源性的克隆ST398。所有菌株中均检测到hla及hld两种溶血素基因,ST239克隆均携带有sea,see,hla,hld以及hlg-2基因,少量菌株还携带有pvl,sed等基因。ST22克隆则携带有包括pvl,seg等8种毒力基因。MRSA菌株对常用抗菌药物呈现高度耐药,MSSA菌株对青霉素、红霉素的耐药率分别为91.3%和69.6%,对其他抗菌药物敏感。结论引起新疆军区总医院血流感染主要MRSA克隆为MRSAST239-t030-SCCmecⅢ,对抗菌药物耐药程度较高;主要MSSA克隆为ST22-t309,携带较多毒力基因,具有高毒力。
OBJECTIVE To investigate the molecular characteristics of clinical Staphylococcus aureus strains isolated from patients with bloodstream infection in Xinjiang Military General Hospital,detect the virulence genes in the strains and analyze the drug resistance to the commonly used antibiotics.METHODS Totally 53 strains of S.aureus were isolated from the patients with bloodstream infection who were treated in Xinjiang Military General Hospital from 2012 to 2016,the drug resistance to methicillin was determined by using polymerase-chain-reaction(PCR),the S.aureus strains were analyzed by means of SCCmec typing,spatyping and multilocus sequence typing(MLST),21 types of virulence genes such as pvl were detected with the use of PCR,the clonal virulence genes that were carried by the strains were analyzed,and the drug resistance of the strains to the commonly used antibiotics was observed.RESULTS Of the 53 strains of S.aureus,30 were methicillin-resistant S.aureus(MRSA),and 23 were methicillin-susceptible S.aureus(MSSA).Totally 4 types of ST,4 types of spaand 3 types of SCCmec were detected in the 30 strains of MRSA,MRSA-ST239-t030-SCCmecⅢ was the predominant prevalent clone,accounting for 90.0%(27/30),meanwhile,3 strains of community-acquired MRSA were detected.Totally10 types of ST and 14 types of spa were detected in the 23 strains of MSSA,ST22-t309 was the prevalent clone,and the animal-derived clone ST398 was detected om 4 strains.All of strains were detected to carry with two types of hemolysin genes hla and hld,the ST239 clone carried with the sea,see,hla,hld and hlg-2 genes,and few strains carried with pvl and sedgenes.ST22 clone strains carried with 8 types of virulence genes,including pvl and seg.The MRSA strains were highly resistant to the commonly used antibiotics,the drug resistance rates of the MSSA strains to penicillin and erythromycin were 91.3%and 69.6%,respectively,and the strains were highly susceptible to other antibiotics.CONCLUSION The MRSA-ST239-t030-SCCmecⅢis dominant among the MRSA clones causing the bloodstream infection in Xinjiang Military General Hospital and is highly resistant to antibiotics.The ST22-t309 is the predominant MSSA clone and carries with multiple virulence genes,which is highly virulent.
OBJECTIVE To investigate the molecular characteristics of clinical Staphylococcus aureus strains isolated from patients with bloodstream infection in Xinjiang Military General Hospital,detect the virulence genes in the strains and analyze the drug resistance to the commonly used antibiotics.METHODS Totally 53 strains of S.aureus were isolated from the patients with bloodstream infection who were treated in Xinjiang Military General Hospital from 2012 to 2016,the drug resistance to methicillin was determined by using polymerase-chain-reaction(PCR),the S.aureus strains were analyzed by means of SCCmec typing,spatyping and multilocus sequence typing(MLST),21 types of virulence genes such as pvl were detected with the use of PCR,the clonal virulence genes that were carried by the strains were analyzed,and the drug resistance of the strains to the commonly used antibiotics was observed.RESULTS Of the 53 strains of S.aureus,30 were methicillin-resistant S.aureus(MRSA),and 23 were methicillin-susceptible S.aureus(MSSA).Totally 4 types of ST,4 types of spaand 3 types of SCCmec were detected in the 30 strains of MRSA,MRSA-ST239-t030-SCCmecⅢ was the predominant prevalent clone,accounting for 90.0%(27/30),meanwhile,3 strains of community-acquired MRSA were detected.Totally10 types of ST and 14 types of spa were detected in the 23 strains of MSSA,ST22-t309 was the prevalent clone,and the animal-derived clone ST398 was detected om 4 strains.All of strains were detected to carry with two types of hemolysin genes hla and hld,the ST239 clone carried with the sea,see,hla,hld and hlg-2 genes,and few strains carried with pvl and sedgenes.ST22 clone strains carried with 8 types of virulence genes,including pvl and seg.The MRSA strains were highly resistant to the commonly used antibiotics,the drug resistance rates of the MSSA strains to penicillin and erythromycin were 91.3%and 69.6%,respectively,and the strains were highly susceptible to other antibiotics.CONCLUSION The MRSA-ST239-t030-SCCmecⅢis dominant among the MRSA clones causing the bloodstream infection in Xinjiang Military General Hospital and is highly resistant to antibiotics.The ST22-t309 is the predominant MSSA clone and carries with multiple virulence genes,which is highly virulent.
引文
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[15]吕媛,李耘,薛峰,等.卫生部全国细菌耐药监测网(Mohnarin)2011-2012年度血流感染细菌耐药监测报告[J].中国临床药理学杂志,2014,30(3):278-288.
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[18]王黎,张文远,卫玮,等.乌鲁木齐地区89例皮肤软组织感染金黄色葡萄球菌分子特征研究[J].中华检验医学杂志,2016,39(6):463-465.
[19]赵春江,王辉,刘颖梅,等.北京四家医院社区获得性甲氧西林耐药金黄色葡萄球菌感染的流行病学调查[J].中华检验医学杂志,2012,35(3):237-242.
[20]顾其芳,张红芝,刘玥,等.金黄色葡萄球菌食品分离株肠毒素基因及耐药性研究[J].中国卫生检验杂志,2018,28(11):1321-1323.
[2]毛镭篥,肖盟,王贺,等.全国多中心细菌耐药监测网中血流感染相关金黄色葡萄球菌的分子流行病学研究[J].中国感染与化疗杂志,2015,15(2):120-125.
[3]杨乐和,余方友,何苏苏,等.血流感染金黄色葡萄球菌的耐药谱分析[J].中华医院感染学杂志,2010,20(18):2869-2871.
[4]Liu C,Chen ZJ,Sun Z,et al.Molecular characteristics and virulence factors in methicillin-susceptible,resistant,and heterogeneous vancomycin-intermediate Staphylococcus aureus from central-southern China[J].J Microbiol Immunol Infect,2015,48(5):490-496.
[5]宋燕,杜昕,田月如,等.六年间血流感染的金黄色葡萄球菌的分子特征分析[J].中华检验医学杂志,2011,34(8):705-711.
[6]杨延成,程航,胡珍,等.乌鲁木齐地区金黄色葡萄球菌分离株的分子分型与耐药性分析[J].第三军医大学学报,2016,38(9):1035-1042.
[7]Kondo Y,Ito T,Ma XX,et al.Combination of multiplex PCRs for staphylococcal cassette chromosome mec type assignment:rapid identification system for mec,ccr,and major differences in junkyard regions[J].Antimicrob Agents Chemother,2007,51(1):264-274.
[8]Zhang K,McClure JA,Elsayed S,et al.Novel multiplex PCRassay for characterization and concomitant subtyping of staphylococcal cassette chromosome mec types I to V in methicillin-resistant Staphylococcus aureus[J].J Clin Microbiol,2005,43(10):5026-5033.
[9]Koreen L,Ramaswamy SV,Graviss EA,et al.spa typing method for discriminating among Staphylococcus aureus isolates:implications for use of a single marker to detect genetic micro-and macrovariation[J].J Clin Microbiol,2004,42(2):792-799.
[10]Harmsen D,Claus H,Witte W,et al.Typing of methicillinresistant Staphylococcus aureus in a university hospital setting by using novel software for spa repeat determination and database management[J].J Clin Microbiol,2003,41(12):5442-5448.
[11]Enright MC,Day NP,Davies CE,et al.Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus[J].J Clin Microbiol,2000,38(3):1008-1015.
[12]Jarraud S,Mougel C,Thioulouse J,et al.Relationships between Staphylococcus aureus genetic background,virulence factors,agr groups(alleles),and human disease[J].Infect Immun,2002,70(2):631-641.
[13]Campbell SJ,Deshmukh HS,Nelson CL,et al.Genotypic characteristics of Staphylococcus aureus isolates from a multinational trial of complicated skin and skin structure infections[J].J Clin Microbiol,2008,46(2):678-684.
[14]Hu Q,Cheng H,Yuan W,et al.Panton-Valentine leukocidin(PVL)-positive health care-associated methicillin-resistant Staphylococcus aureus isolates are associated with skin and soft tissue infections and colonized mainly by infective PVL-encoding bacteriophages[J].J Clin Microbiol,2015,53(1):67-72.
[15]吕媛,李耘,薛峰,等.卫生部全国细菌耐药监测网(Mohnarin)2011-2012年度血流感染细菌耐药监测报告[J].中国临床药理学杂志,2014,30(3):278-288.
[16]Teixeira LA,Resende CA,Ormonde LR,et al.Geographic spread of epidemic multiresistant Staphylococcus aureus clone in Brazil[J].J Clin Microbiol,1995,33(9):2400-2404.
[17]申恩华,王立红,王辉,等.中国15个地区分离的耐甲氧西林金黄色葡萄球菌基因分型研究[J].中华流行病学杂志,2010,31(3):308-311.
[18]王黎,张文远,卫玮,等.乌鲁木齐地区89例皮肤软组织感染金黄色葡萄球菌分子特征研究[J].中华检验医学杂志,2016,39(6):463-465.
[19]赵春江,王辉,刘颖梅,等.北京四家医院社区获得性甲氧西林耐药金黄色葡萄球菌感染的流行病学调查[J].中华检验医学杂志,2012,35(3):237-242.
[20]顾其芳,张红芝,刘玥,等.金黄色葡萄球菌食品分离株肠毒素基因及耐药性研究[J].中国卫生检验杂志,2018,28(11):1321-1323.