淮南市O139霍乱疫情3种快速检测方法的应用比较及菌株相似性分析  

The comparison of three kinds of emergency detection methods applied in O139 cholera epidemic and the analysis of strains homology in Huainan

作　　者：刘江[1] 孙永[2] 余寿杰[1] 刘笑梅[1] 曾凡荣[1] 王晨琨[1] 

机构地区：[1]淮南市疾病预防控制中心微检科,安徽淮南232033 [2]安徽省疾病预防控制中心微检科,安徽合肥230601

出　　处：《中国卫生检验杂志》2015年第1期47-49,共3页Chinese Journal of Health Laboratory Technology

摘　　要：目的通过建立霍乱弧菌的快速检测方法,提高霍乱疫情应急检测的速度和检出率,溯源可疑的传染源。方法将细菌分离鉴定,实时荧光PCR快速检测,胶体金免疫层析快速试纸法同时用于检测霍乱疫情处置期间所采集的患者水样便、外环境水质、食品及水产品,并对其结果进行比较分析;将分离到的O139霍乱弧菌进行脉冲场凝胶(PFGE)电泳分型分析。结果 3种检测方法中实时荧光PCR快速检测和胶体金免疫层析快速试纸法阳性率较高,常规细菌分离鉴定阳性率较低,3种方法各有优势及缺点;PFGE分子分型分析提示本次O139霍乱疫情传染源为水产市场中甲鱼带菌,污染病人食用食物所致。结论荧光PCR快速检测和胶体金免疫层析快速试纸法是菌株分离培养方法的有效补充;PFGE对不同来源的霍乱弧菌遗传同源相关性分析有较好的分辨能力。Objective To establish rapid detection method of Vibrio cholerae,so as to improve the speed and emergency detection rate for the cholera epidemic,and trace the suspected source of infection. Methods To isolate and identify bacteria,as well as conduct rapid detection with real- time PCR. Colloidal gold immune chromatography（ GICA） was simultaneously used for the detection of collected watery stool,external environment water quality,food and aquatic products during dealing with cholera,and the results were compared; The separated Vibrio cholerae O139 was genotyped by pulsed- field gel electrophoresis（ PFGE）. Results In the three kinds of detection methods,the positive detection rate of real- time PCR and GICA was higher,that of normal bacteria isolation was lower,but these three methods had their own advantages and drawbacks; Pulsed- field condensate glue electrophoresis（ PFGE） molecular typing analysis suggested the source of this infection for the O139 cholera epidemic came from snapper fish. Conclusion Fluorescence PCR for rapid detection and GICA were effective supplement for strains isolation and culture method; PFGE had the ability to distinguish good genetic homology of Vibrio cholera from different sources.

关 键 词：O139霍乱弧菌 检测方法 细菌分离鉴定 聚合酶链式反应 胶体金免疫层析 脉冲场凝胶电泳 

分 类 号：R516.5[医药卫生—内科学]