CAJ | 학술논문

目的了解深圳地区副溶血弧菌主要血清型和分子分型以及毒力因子分布,分析新O3:K6型克隆群与国际流行株的进化关系.方法对2002-2008年深圳地区1005株副溶血弧菌临床分离株进行血清型分型；采用荧光PCR方法对68种不同血清型的28 1株副溶血弧菌进行tlh、toxR、tdh和trh毒力基因检测；对281株菌株进行orf8检测；采用脉冲场凝胶电泳(PFGE)对不同分离时间的5种主要血清型菌株进行分子分型,根据有代表性的分子分型图谱选取82株副溶血弧菌菌株进行GS-PCR检测,并选取60株副溶血弧菌菌株进行toxRS基因测序；进而对41株O3∶K6和O1∶K25副溶血弧菌菌株进行多位点序列分型(MLST)分析,采用eBURST软件分析进化关系.结果 1005株菌共得到79种不同的血清型,主要血清型为O3∶K6、O4∶K8、O1∶K25、O1∶KUT、O4∶K68、O1∶K56和O9∶K44,所占比例分别为57.9％、8.16％、5.27％、5.87％、1.39％、1.39％和0.99％.243株为tlh+、toxR+、tdh+、trh-,37株为tlh+、toxR+、tdh-和trh-,1株tlh+、toxR+、tdh+和trh+.35株O3∶K6型菌株的MLST序列类型为ST3,是同源复合体CC3,O4∶K8型副溶血弧菌的序列类型为ST189,无同源复合体.结论深圳地区主要流行的副溶血弧菌为tdh+、trh-的O3∶K6、O4∶K8和O1∶K25菌株,2006年后出现O1∶K56、O9∶K44、O3∶K29、O4∶K9新的血清型；其中O3∶K6血清型属于新的O3∶K6克隆群,与其他国家流行的副溶血弧菌属于同一克隆群,同时也存在新、旧克隆群.血清的多样性和新旧克隆群的存在是造成深圳地区副溶血弧菌腹泻病高发态势的主要原因之一.
목적 료해심수지구부용혈호균주요혈청형화분자분형이급독력인자분포,분석신O3:K6형극륭군여국제류행주적진화관계.방법 대2002-2008년심수지구1005주부용혈호균림상분리주진행혈청형분형；채용형광PCR방법대68충불동혈청형적28 1주부용혈호균진행tlh、toxR、tdh화trh독력기인검측；대281주균주진행orf8검측；채용맥충장응효전영(PFGE)대불동분리시간적5충주요혈청형균주진행분자분형,근거유대표성적분자분형도보선취82주부용혈호균균주진행GS-PCR검측,병선취60주부용혈호균균주진행toxRS기인측서；진이대41주O3∶K6화O1∶K25부용혈호균균주진행다위점서렬분형(MLST)분석,채용eBURST연건분석진화관계.결과 1005주균공득도79충불동적혈청형,주요혈청형위O3∶K6、O4∶K8、O1∶K25、O1∶KUT、O4∶K68、O1∶K56화O9∶K44,소점비례분별위57.9％、8.16％、5.27％、5.87％、1.39％、1.39％화0.99％.243주위tlh+、toxR+、tdh+、trh-,37주위tlh+、toxR+、tdh-화trh-,1주tlh+、toxR+、tdh+화trh+.35주O3∶K6형균주적MLST서렬류형위ST3,시동원복합체CC3,O4∶K8형부용혈호균적서렬류형위ST189,무동원복합체.결론 심수지구주요류행적부용혈호균위tdh+、trh-적O3∶K6、O4∶K8화O1∶K25균주,2006년후출현O1∶K56、O9∶K44、O3∶K29、O4∶K9신적혈청형；기중O3∶K6혈청형속우신적O3∶K6극륭군,여기타국가류행적부용혈호균속우동일극륭군,동시야존재신、구극륭군.혈청적다양성화신구극륭군적존재시조성심수지구부용혈호균복사병고발태세적주요원인지일.
Objective To determine the occurrence and distribution of specific clones of pathogenic Vibrio parahaemolyticas (VP) isolated in Shenzhen and to assess the relationship between serotype O3 ∶K6 and the globally distributed pandemic clone.Methods A total of 1005 VPs isolated from diarrhea patients in 2002-2008 were sero-typed.Real-time PCR was used to detect the virulence genes tlh,toxR,tdh,trh and orf8 in 281 isolates from 68 different serotypes.The main serotypes were typed by pulsed field gel electrophoresis (PFGE).Strains with dominant serotypes and PFGE pattems were assayed by GS-PCR and toxRS sequencing for the identification of pandemic clone.Multilocus sequence typing (MLST) analysis was reserved for exemplary 41 O3 ∶ K6 and O1 ∶ K25 isolates.Results Seventy-nine serotypes were observed among the 1005 isolates,including O3 ∶ K6 (57.9％),O4 ∶ K8 (8.16％),O1 ∶ KUT (5.87％),O1 ∶ K25 (5.27％),O4 ∶ K68(1.39％),O1 ∶ K56(1.39％) and O9 ∶ K44 (0.99％).Most of the strains (99.36％) showed PCR positive to tlh,toxR,and tdh but eleven strains were tdh negative.MLST showed that all the 36 O3 ∶ K6 isolates belonged to ST3 and all the 5 O4∶K8 strains were ST189.These results matched the description of the pandemic VP clone.Conclusion A recognizable burden of diarrheal illness caused by VP had been seen in Shenzhen.Results from serotyping indicated that although there exsiting a large variety of diversities,the dominant serotype appeared to be O3 ∶ K6.VP isolates identified in Shenzhen mainly showed as tdh positive but trh negative,in consistent with the current pandemic O3 ∶ K6 clone.The pandemic O3 ∶ K6 clone did appear to co-exist with other clones of O3 ∶ K6,as well as O4 ∶ K8,O1 ∶ K25.Potential outbreak of VP could be monitored through the laboratory-based surveillance programs,suggesting that the strategies related to prevention and control of VP should be prioritized in Shenzhen.