[1]庞毅恒 杜 悦 王 红.连续传代对副溶血性弧菌分子分型的影响[J].大众科技,2019,21(02):43-46.
　Effects of Continuous Passage on Molecular Typing of Vibrio Parahaemolyticus[J].Popular Science & Technology,2019,21(02):43-46.
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连续传代对副溶血性弧菌分子分型的影响()
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《大众科技》[ISSN:1008-1151/CN:45-1235/N]

卷:
21
期数:
2019年02
页码:
43-46
栏目:
医药与卫生
出版日期:
2019-02-20

文章信息/Info

Title:
Effects of Continuous Passage on Molecular Typing of Vibrio Parahaemolyticus
作者:
庞毅恒1 杜 悦2 王 红2
（1.广西医科大学第二附属医院，广西 南宁 530007； 2.广西壮族自治区疾病预防控制中心，广西 南宁 530028）
关键词:
副溶血性弧菌; 脉冲场凝胶电泳; 多位点序列分析; 传代培养
Keywords:
Vibrio parahaemolyticus;  pulsed field gel electrophoresis;  multilocus sequence typing;  passage
文献标志码:
A
摘要:
目的：评价副溶血性弧菌经多次传代培养后，对脉冲场凝胶电泳所获得的指纹图谱存在的影响。方法：将从腹泻患者粪便分离到的两株副溶血性弧菌复苏，进行生化鉴定，血清诊断等表型分析；用3%NaCl 碱性蛋白胨水进行连续传代35代，每代次菌株用含3%NaCl TSA 划线培养，收集3%NaCl TSA 过夜培养物进行多位点序列分析，用Not I 限制性内酶切进行脉冲场凝胶电泳分型。通过生物信息学方法模拟PFGE 分析标准株Not I 酶切片段大小。结果：复苏的菌株确定为副溶血性弧菌，经连续传35 代后，各代次的基因指纹图谱总体上呈现较稳定的型别，但在个别代次中会出现细微差异而后会恢复；传代对多位点序列分析没有影响。结论：传代对某些菌型的副溶血性弧菌的多位点序列分析和脉冲场凝胶电泳基因指纹图谱影响不大；脉冲场凝胶电泳可以对副溶血性弧菌菌株进行溯源。
Abstract:
Objective: To evaluate the effect of Vibrio parahaemolyticus on the fingerprints obtained by pulsed field gel electrophoresis after multiple passages. Methods: The two tested strains of Vibrio parahaemolyticus isolated from the feces of diarrhea patients were resuscitated for biochemical identification, serodiagnosis and other phenotypic analysis, and then were sub-cultivated for 35 generations through 3% NaCl alkaline peptone solution. Each generation of strains was collected from 3% NaCl TSA overnight culture for multilocus sequence typing and pulsed field gel electrophoresis by Not I restriction endonuclease digestion. The DNA fingerprints of the standard strain by Not I digestion was simulated by bioinformatics method. Results: The DNA fingerprints of these strains of pulsed field gel electrophoresis in each generation showed roughly a stable type, though there were subtle differences in individual generations. The multilocus sequence typing of strains showed no difference in each generation. Conclusion: The continuous-repeated passage of Vibrio parahaemolyticus has little effect on the multi-locus sequence analysis and pulsed field gel electrophoresis gene fingerprinting. It was a pretty technology for pulsed field gel electrophoresis to trace the source strain of Vibrio parahaemolyticus in foodborne diseases.

参考文献/References:

【参考文献】 [1] 郑文龙,王卓,马洁,等. 副溶血性弧菌病原学和分子流行病学流行特征研究进展[J].疾病监测,2015,30(4):337-341. [2] 黄彦,唐振柱,李秀桂,等.我国食物中毒副溶血性弧菌流行株血清型及分子分型现况[J].职业与健康,2013,29(8):998- 1000. [3] Martinezurtaza J,Powell A,Jansa J,et al.Epidemiological investigation of a foodborne outbreak in Spain associated with U.S.West Coast genotypes of Vibrio parahaemolyticus [J].Springerplus,2016,5(1):87. [4] 王红,赵鹏,杜悦,等.多菌型副溶血性弧菌共感染引起集中事件调查分析[J].疾病监测,2018,33(4):312-315. [5] 李伟,崔志刚,阚飙,等.中国细菌性传染病分子分型实验室监测网络-PulseNet China[J].疾病监测,2011,26(1):1-4. [6] Ward L N,Bej A K.Detection of Vibrio parahaemolyticus in shellfish by use of multiplexed real-time PCR with TaqMan fluorescent probes[J].Applied & Environmental Microbiology, 2006,72(3):2031-2042. [7] 吴青,韩海红,余东敏,等.北京市水产品污染与感染病例中副溶血性弧菌血清型和毒力基因型的比较研究[J].中国食品卫生杂志,2015,27(4):363-367.

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备注/Memo

备注/Memo:
【收稿日期】2018-12-13 【基金项目】广西医疗卫生适宜技术开发与推广应用项目（S2017065）；广西自然科学基金面上项目（2017GXNSFAA198270）；广西高校中青年教师基础能力提升项目（2017KY0123）。 【作者简介】庞毅恒（1978－），女，广西南宁人，广西医科大学第二附属医院教务部副主任医师，从事医疗卫生工作。【通信作者】王红，广西壮族自治区疾病预防控制中心副主任技师。
更新日期/Last Update: 2019-04-02