中国感染与化疗杂志
中國感染與化療雜誌
중국감염여화료잡지
CHINESE JOURNAL OF INFECTION AND CHEMOTHERAPY
2015年
3期
222-229
,共8页
蒋浩琴%陈明亮%李天铭%朱元军%刘红%曾玫%李敏
蔣浩琴%陳明亮%李天銘%硃元軍%劉紅%曾玫%李敏
장호금%진명량%리천명%주원군%류홍%증매%리민
A群链球菌%药敏试验%多位点序列分型%emm分型%脉冲场凝胶电泳%生物膜%毒力基因
A群鏈毬菌%藥敏試驗%多位點序列分型%emm分型%脈遲場凝膠電泳%生物膜%毒力基因
A군련구균%약민시험%다위점서렬분형%emm분형%맥충장응효전영%생물막%독력기인
Streptococcus pyogenes%antimicrobial susceptibility testing%multilocus sequence typing%emm typing%pulsed field gel electrophoresis%biofilm%virulence factor
目的：研究和比较上海成人和儿童患者分离的A群链球菌（GAS）耐药、克隆分型、emm分型、生物膜形成及毒力因子携带等分子特征，为感染控制及治疗提供临床流行病学信息。方法39株成人（13株）和儿童（26株）患者分离的GAS ，用K‐B纸片法测定对9种常用抗菌药物的敏感性；采用多位点序列分型（MLST）方法进行克隆分型；采用编码M蛋白的emm基因序列分析进行基因分型；采用脉冲场凝胶电泳（PFGE）方法分析不同 emm型菌株的基因组特征；采用半定量生物膜形成试验分析其生物膜形成。聚合酶链反应（PCR）方法检测20个包括超抗原在内的GAS主要毒力基因。结果39株GAS主要基因型为emm12‐ST36（64．1％），emm1‐ST28（17．9％）；成人和儿童均以emm12‐ST36为主。儿童菌株对红霉素和克林霉素的耐药率显著高于成人（P＝0．0087）。相同emm分型‐克隆分型菌株在PFGE脉冲场带型中呈现高度聚集。生物膜形成与 emm1型菌株明显相关（P＝0．005），与红霉素和克林霉素耐药密切相关（P＝0．0003），儿童菌株的生物膜形成强于成人菌株（P＜0．0001）。毒力基因 speG、speB、sdaB、Mac全部阳性；speA、speJ、spd3与 emm1型菌株有明显相关（分别为 P＜0．0001、P＝0．0055、P＜0．0001）；speI、sic与emm12型菌株有明显相关（均为 P＜0．0001）；speH、ssa在emm12和 emm1型菌株中有明显分布差异（分别为 P＝0．0364、P＝0．0258）；20个毒力基因在成人和儿童emm12型菌株中均没有明显分布差异（均为 P＞0．05）。结论 emm分型与克隆分型、PFGE、毒力基因有很好相关性。成人和儿童菌株在耐药情况、生物膜形成中均存在差异。特定 emm型菌株与抗生素耐药和致病力密切相关，为感染控制提供依据。重要毒力基因将是未来研制新型疫苗降低GAS感染的新型靶标。
目的：研究和比較上海成人和兒童患者分離的A群鏈毬菌（GAS）耐藥、剋隆分型、emm分型、生物膜形成及毒力因子攜帶等分子特徵，為感染控製及治療提供臨床流行病學信息。方法39株成人（13株）和兒童（26株）患者分離的GAS ，用K‐B紙片法測定對9種常用抗菌藥物的敏感性；採用多位點序列分型（MLST）方法進行剋隆分型；採用編碼M蛋白的emm基因序列分析進行基因分型；採用脈遲場凝膠電泳（PFGE）方法分析不同 emm型菌株的基因組特徵；採用半定量生物膜形成試驗分析其生物膜形成。聚閤酶鏈反應（PCR）方法檢測20箇包括超抗原在內的GAS主要毒力基因。結果39株GAS主要基因型為emm12‐ST36（64．1％），emm1‐ST28（17．9％）；成人和兒童均以emm12‐ST36為主。兒童菌株對紅黴素和剋林黴素的耐藥率顯著高于成人（P＝0．0087）。相同emm分型‐剋隆分型菌株在PFGE脈遲場帶型中呈現高度聚集。生物膜形成與 emm1型菌株明顯相關（P＝0．005），與紅黴素和剋林黴素耐藥密切相關（P＝0．0003），兒童菌株的生物膜形成彊于成人菌株（P＜0．0001）。毒力基因 speG、speB、sdaB、Mac全部暘性；speA、speJ、spd3與 emm1型菌株有明顯相關（分彆為 P＜0．0001、P＝0．0055、P＜0．0001）；speI、sic與emm12型菌株有明顯相關（均為 P＜0．0001）；speH、ssa在emm12和 emm1型菌株中有明顯分佈差異（分彆為 P＝0．0364、P＝0．0258）；20箇毒力基因在成人和兒童emm12型菌株中均沒有明顯分佈差異（均為 P＞0．05）。結論 emm分型與剋隆分型、PFGE、毒力基因有很好相關性。成人和兒童菌株在耐藥情況、生物膜形成中均存在差異。特定 emm型菌株與抗生素耐藥和緻病力密切相關，為感染控製提供依據。重要毒力基因將是未來研製新型疫苗降低GAS感染的新型靶標。
목적：연구화비교상해성인화인동환자분리적A군련구균（GAS）내약、극륭분형、emm분형、생물막형성급독력인자휴대등분자특정，위감염공제급치료제공림상류행병학신식。방법39주성인（13주）화인동（26주）환자분리적GAS ，용K‐B지편법측정대9충상용항균약물적민감성；채용다위점서렬분형（MLST）방법진행극륭분형；채용편마M단백적emm기인서렬분석진행기인분형；채용맥충장응효전영（PFGE）방법분석불동 emm형균주적기인조특정；채용반정량생물막형성시험분석기생물막형성。취합매련반응（PCR）방법검측20개포괄초항원재내적GAS주요독력기인。결과39주GAS주요기인형위emm12‐ST36（64．1％），emm1‐ST28（17．9％）；성인화인동균이emm12‐ST36위주。인동균주대홍매소화극림매소적내약솔현저고우성인（P＝0．0087）。상동emm분형‐극륭분형균주재PFGE맥충장대형중정현고도취집。생물막형성여 emm1형균주명현상관（P＝0．005），여홍매소화극림매소내약밀절상관（P＝0．0003），인동균주적생물막형성강우성인균주（P＜0．0001）。독력기인 speG、speB、sdaB、Mac전부양성；speA、speJ、spd3여 emm1형균주유명현상관（분별위 P＜0．0001、P＝0．0055、P＜0．0001）；speI、sic여emm12형균주유명현상관（균위 P＜0．0001）；speH、ssa재emm12화 emm1형균주중유명현분포차이（분별위 P＝0．0364、P＝0．0258）；20개독력기인재성인화인동emm12형균주중균몰유명현분포차이（균위 P＞0．05）。결론 emm분형여극륭분형、PFGE、독력기인유흔호상관성。성인화인동균주재내약정황、생물막형성중균존재차이。특정 emm형균주여항생소내약화치병력밀절상관，위감염공제제공의거。중요독력기인장시미래연제신형역묘강저GAS감염적신형파표。
Objective To investigate and compare the molecular epidemiological characteristics of Streptococcus pyogenes isolates from Shanghai adult and pediatric patients in terms of antimicrobial susceptibility ,clone type ,emm type ,biofilm formation and virulence for better infection control and treatment .Methods Thirty‐nine nonduplicate clinical isolates of S . pyogenes from adult and pediatric patients were analyzed by determining the susceptibility to antimicrobial agents by Kirby‐Bauer method;clonal typing by multilocus sequence typing ( MLST ); genotyping by emm gene sequence analysis ,which encoding M protein;genomic characteristics of different emm type strains by pulsed field gel electrophoresis (PFGE );and biofilm formation by semi‐quantitative biofilm formation test . Twenty main virulence genes of S .pyogenes ,including 12 superantigen genes and 8 other key genes were detected by PCR and gel electrophoresis . Results A total of 39 nonduplicate S .pyogenes isolates were analyzed .The most common genotype was emm 12‐ST36 (64 .1% ) and emm 1‐ST28 (17 .9% ) .Isolates from adult and pediatric patients had the same dominant genotype , emm 12‐ST36 . The isolates from children showed significantly higher resistance rate to erythromycin and clindamycin than those from adult patients (P<0 .000 1) .Particular emm type and clone type were frequently identified in the same PFGE cluster .Statistical analysis showed that biofilm formation was significantly associated with emm type 1 (P=0 .005) and erythromycin/clindamycin resistance (P=0 .000 3) .The strains from children showed higher biofilm formation than those from adult patients (P<0 .000 1) .We found that virulence genes speA ,speJ and spd3 were significantly associated with emm type 1 (P<0 .000 1 ,P=0 .005 5 ,P<0 .000 1) ,while speI and sic were significantly associated with emm type 12 (both P<0 .000 1) .We also found that the prevalence of speC ,speH ,ssa , smeZ ,and sdaD genes was significantly different between emm type 12 and emm type 1 (P= 0 .023 8 , P< 0 .000 1 , P<0.0001,P= 0.0003,and P= 0.0068,respectively).TheprevalenceofvirulencegenesspeH,smeZandsdaDwas significantly different between the emm type 12 strains from children and those from adults (all P< 0 .000 1) .Conclusions There is a strong agreement between emm type ,clone type ,virulence genes and the clusters defined by PFGE profiling of S . pyogenes .S .pyogenes isolates from adult and pediatric patients are different in terms of antibiotic resistance and biofilm formation .Certain emm type is significantly associated with antibiotic resistance and virulence ,which is useful for infection control .Dominant virulence genes may be the potential target for developing new vaccine to reduce S .pyogenes infection in the future .
