疑似霍乱弧菌鉴定及分子分型检测  

Characterization and molecular typing of suspected Vibrio cholerae isolates

作　　者：陈琛[1] 王中强[1] 柳楠[1] 刘雯静[1] 李婧[1] 薛文仲[1] 张伶[1] 杜昕颖[1] 邱少富[1] 王勇[1] 黄留玉[1] 李承毅[1] 宋宏彬[1] 

机构地区：[1]中国人民解放军疾病预防控制所应急大队,北京100071

出　　处：《中国公共卫生》2011年第1期57-58,共2页Chinese Journal of Public Health

基　　金：国家科技重大专项(2008ZX10004-0082009ZX10004-2052009ZX10004-315);国家自然科学基金(81001267/H2609)

摘　　要：目的对北京市某医院肠道门诊2株疑似霍乱弧菌进行鉴定,检测其主要毒力基因,并分析其基因型别特征。方法利用细菌生化鉴定条(API 20 E)进行生化鉴定,玻片凝集法确定其血清型别,应用PCR及测序技术分析其16 S rRNA基因;PCR检测其8个主要毒力基因,脉冲场凝胶电泳(PFGE)分析其基因型别。结果经鉴定,该2株疑似霍乱菌株均为非O1非139群霍乱弧菌,经16 SrRNA分析与美国国家生物技术信息中心数据库中霍乱弧菌相似性达100%,均包含毒力基因hylA、ompW和toxR,PFGE结果显示2株菌具有完全不同的带型。结论 2株疑似霍乱弧菌为非O1非139群霍乱弧菌,其致病因素与(hylA)、(ompW)、(toxR)毒力基因有关,并且2株菌遗传关系较远。Objective To identify two suspected Vibrio cholerae isolates from patients with intestinal diseases diagnosed in a hospital in Beijing city and to detect their main virulence genes and analyze their genotypes.Methods The suspected V.cholerae isolates were identified with API 20E strips,serotype and 16S rRNA analysis.PCR was used to detect the distribution of the eight major virulence genes,and the genetic association was measured with pulsed-field gel electrophoresis（PFGE）.Results The two suspected V.cholerae isolates were confirmed as non-O1/non-O139 V.cholerae.16S rRNA analysis showed that the two isolates was 100% homologous to those from NCBI database.Virulence genes hylA,ompW and toxR were detected.The two strains showed different banding patterns with PFGE.Conclusion The two suspected strains were identified to be non-O1/non-O139 V.cholera and the pathogenic factors may be related to the virulence genes hylA,ompW and toxR.The two strains were distant in evolutionary relationship.The result indicates that different types of non-O1/non-O139 V.cholera exist in the city.

关 键 词：霍乱弧菌 毒力基因 脉冲场凝胶电泳 

分 类 号：Q939.121[生物学—微生物学]