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目的:对食品从业人员粪便标本进行霍乱弧菌监测,对分离到的霍乱弧菌进行病原学鉴定,为霍乱防治提供信息。方法:按照W S289-2008《霍乱诊断标准》和《霍乱防治手册》第五版,分离霍乱弧菌,并确定所分离的霍乱弧菌血清型和生化反应特性;采用噬菌体生物分型区别流行和非流行株及不同菌型;采用PCR检测霍乱弧菌分离菌株ctxA和tc-pA毒力基因携带情况;采用脉冲场凝胶电泳(Pu lsed field gelelectrophoresis,PFGE)进行分子分型;采用K-B纸片法检测分离株对14种抗生素的敏感性。结果:从辖区同一家餐饮店不同工种的食品从业人员肛拭子中检出3株O1群霍乱弧菌小川血清型。3株菌噬菌体生物分型均为15 e,属埃尔托生物型非流行菌株,霍乱弧菌毒力基因PCR检测结果为ctxA阴性,tcpA阳性,3株菌PFGE分型结果一致。结论:检出的3株O1群霍乱弧菌埃尔托生物型虽然为非流行菌株,但从同一家餐饮店在同一时间段同时检出,杭州市尚属首例,应引起密切关注。传统的细菌系统鉴定结合分子分型方法,为霍乱弧菌的快速检测和流行病学溯源提供了技术支持。
OBJECTIVE: To monitor Vibrio cholerae in the stool samples of food workers and to identify the isolated Vibrio cholerae and provide information for prevention and control of cholera. Methods: Vibrio cholerae were isolated and the serotypes and biochemical reactions of V. cholerae were isolated according to the Diagnostic Criteria of Cholera and the fifth edition of Cholera Prevention and Treatment Manual. The phage biomarker was used to distinguish the epidemic from Non-epidemic strains and different strains. PCR was used to detect virulence genes of ctxA and tc-pA in V. cholerae isolates. Molecular typing was performed by pulsed field gel electrophoresis (PFGE) Method to test the sensitivity of isolates to 14 antibiotics. Results: Three strains of Vibrio cholera O1 O1 serogroup O1 were detected from the anal swabs of food practitioners of different types in the same restaurant in the area. The three strains of phage were 15 e, which belongs to the Elto biotype non-epidemic strains. The PCR results of Vibrio cholerae virulence gene were ctxA negative and tcpA positive, and the results of PFGE typing of the three strains were consistent. Conclusion: Although the Elto biotypes of the three strains of Vibrio cholerae O1 were detected as non-epidemic strains, Hangzhou was the first case from the same restaurant at the same time period, which should be closely concerned. Traditional bacterial system identification combined with molecular typing methods provide technical support for the rapid detection and epidemiological tracing of V. cholerae.