副溶血弧菌临床分离株毒力特征与多位点序列分型研究  

Virulence characteristics and multilocus sequence type of Vibrio parahaemolyticus isolated from clinic

作　　者：范艳艳[1] 朱敏[1] 尚欣荣[1] 王玫[1] 黄艳飞[1] 顾海彤[1] 鲁辛辛[1] 

机构地区：[1]首都医科大学附属北京同仁医院检验科,100730

出　　处：《中华检验医学杂志》2013年第6期548-552,共5页Chinese Journal of Laboratory Medicine

摘　　要：目的探讨副溶血弧菌（VP）临床分离株的毒力特征及分子分型特征。方法收集2009至2011年北京同仁医院肠道门诊腹泻患者分离的152株VP，采用PCR技术检测溶血毒素基因（tdh、trh）、两个Ⅲ型分泌系统2（T3SS2α、T3ss2β）及03：K6克隆群系统性毒力基因（toxRS／new、orf8）。采用多位点序列分型（MLST）技术对菌株进行分子分型研究。结果152株VPtdh阳性率为96％（146／152），trh阳性率为2％（3／152），T3SS2阳性率为100％（152／152）。实验中分离到大流行株（tdh＋、toxRS／new＋、trh-）107株，致病株（tdh＋或／和trh＋）38株，非致病株（tdh-且trh-）6株。其中非致病株均含大流行株系统性毒力基因oft8和toxRS／new，仅1株致病株含068基因；另发现一分离株含全部毒力基因。MLST分型将152株VP分为16个序列型（ST型），其中ST3为优势序列型，包含113株菌，并发现4种新ST型。结论本研究副溶血弧菌90％以上携带tdh，以ST3型为主要流行株，通过毒力基因检测和MLST研究可快速追踪流行株致病性和分子进化信息。Objective To study on virulence characteristics and multilocus sequence type of Vibrio parahaemolyticus isolated from clinic in Beijing Tongren hospital. Methods Total 152 strains of Vibrio parahaemolyticus isolates were collected from diarrheal patients of outpatients in Beijing Tongren Hospital, Capital Medical University from 2009 to 2011. PCR was used to detect hemolysin gene thermo stable direct themolysin gene （tdh）,TDH-related hemolysin gene （trh）, type Ⅲ secretion system 2 （T3SS2α ,T3SS2β） and systematic functional gene （toxRS/new, ofr8 ） for pandemic O3：K6 clone and its derivatives. The genetic features of these strains were determined by multiloeus sequence typing （MLST）. Results 96% （ 146/152 ） VP harbored tdh gene, 2% （3/152） VP harbored trh gene and 100% （152/152） VP harbored T3SS2 gene. In this study there were 107 pandemic strains （ both tdh and toxRS/new positive and trh negative）, 38 pathogenic strains （tdh positive and/or trh positive） and 6 nonpathogenic strains （both tdh and trh negative）. All nonpathogenic strains harbored systematic functional gene （toxRS/new, orf8 ）. Only one pathogenic strains harbored off8 gene. One clone harbored all virulence gene. In this study there were 16 sequence types, and ST3 is the pandemic sequence type, including 113 strains, and four new sequence types were found. Conclusions In this study more than 90% Vibrio haemolyticus harbored tdh gene and ST3 was the pandemic sequence type in Beijing. One can get bacterial pathogenic charateristic and population genetics information by virulence gene testing and MLST.

关 键 词：弧菌 副溶血性 毒力 多位点序列分型 流行病学 分子 

分 类 号：R379.4[医药卫生—病原生物学]