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目的了解副溶血弧菌临床分离株的毒力基因、血清型及基因分型特征,探讨多位点序列分型(MLST)分型技术对其同源性分析的应用价值。方法收集深圳地区2007-2014年食物中毒及医院散发病例来源的68株副溶血弧菌菌株,对其进行血清型、毒力基因分析。选择副溶血弧菌MLST数据库提供的7个管家基因,PCR扩增测序后采用Bio Edit、e BURST等软件,对本次研究的68株副溶血弧菌进行MLST同源性分析。结果 68株副溶血弧菌共得到7种不同的血清型,优势血清型为O3∶K6(47/68),其余血清型分别为O4∶K8、O5∶K68、O4∶K13等。副溶血弧菌临床分离株94%为tdh+trh-。MIST分型将68株副溶血弧菌分为5个序列型(ST型),ST3为优势序列型(55/68),其余4种ST型中的ST189与ST345构成一个同源复合体CC345,且ST189为ST345的单位点变异型。结论深圳地区副溶血弧菌所致的腹泻病暴发疫情,其流行菌株ST型别多样化,疾病控制中需警惕非主流ST型别引起暴发可能。
Objective To understand the virulence genes, serotypes and genotyping characteristics of clinical isolates of Vibrio parahaemolyticus, and to explore the value of multi-locus sequencing (MLST) typing in homology analysis. Methods A total of 68 strains of Vibrio parahaemolyticus from food poisoning and hospital-issued cases from 2007 to 2014 in Shenzhen were collected and analyzed for their serotypes and virulence genes. Seven housekeeping genes were selected from the MLST database of Vibrio parahaemolyticus. The MLST homology analysis of 68 strains of Vibrio parahaemolyticus in this study was performed by using software such as Bio Edit and e BURST after PCR amplification and sequencing. Results A total of seven strains of Vibrio parahaemolyticus were obtained. The predominant serotypes were O3: K6 (47/68). The remaining serotypes were O4: K8, O5: K68, O4: K13 and so on. 94% of clinical isolates of Vibrio parahaemolyticus were tdh + trh-. Sixty-eight strains of Vibrio parahaemolyticus were divided into five sequence types (ST type) and ST3 (55/68), and ST189 and ST345 of the other four ST types constituted a homologous complex CC345. And ST189 is a single-site variant of ST345. Conclusion The outbreak of diarrheal disease caused by Vibrio parahaemolyticus in Shenzhen area is diversified in the ST type of its epidemic strain. It is necessary to be alert to the possibility of outbreak caused by non-mainstream ST type in disease control.