不同来源的霍乱弧菌毒力特征的质谱学分析  

Virulence Characteristics Application of Vibrio cholera from Different Source by Microbial Mass Spectrometry

作　　者：屠博文[1] 薛银刚 姚萍[1] 王楠 黎俊宏[1] 唐宏兵[1] 杜强[1] TU Bowen;XUE Yingang;YAO Ping;WAND Nan;LI Junhong;TANG Hongbing;DU Qiang(Changzhou Center of Disease Prevention and Control,Changzhou 213022,China;School of Environmental and Safety Engineering,Changzhou University,Changzhou 213003,China;Medical device department of market supervision administration of Taizhou pharmaceutical high-tech zone,Taizhou 225300,China)

机构地区：[1]常州市疾病预防控制中心,江苏常州213022 [2]常州大学环境与安全工程学院,江苏常州213003 [3]泰州医药高新区市场监督管理局医疗器械处,江苏泰州225300

出　　处：《食品工业科技》2021年第6期130-136,150,共8页Science and Technology of Food Industry

基　　金：江苏省卫生健康委科研课题面上项目(H2018098);常州市重大科技支撑计划(社会发展)(CE20185048和CE 20175022);江苏省预防医学会课题(Y2018023);常州市应用基础研究计划(CJ20190071)。

摘　　要：目的:分析本地区食品风险监测样品中霍乱弧菌的分布,评价使用VITEK-MS微生物质谱进行霍乱弧菌快速鉴定的优劣。方法:从淡水产品和病人分离获得霍乱弧菌,分别进行VITEK-2生化鉴定和VITEK-MS质谱鉴定。血清凝集和胶体金显色法检测霍乱弧菌血清型,PCR检测ctx毒力基因。通过全基因组测序分析霍乱弧菌毒力基因类型。结果:经快速鉴定检出23株霍乱弧菌,17株为非O1/O139血清型,6株为O139型并且携带ctx毒力基因。VTIEK-MS微生物质谱检出的霍乱弧菌具有典型的指纹图谱特征,O139型霍乱弧菌具有典型的特有峰和缺失峰特征。全基因组测序显示O139血清型具ctxAB、ace和tcp毒力岛等毒力基因,而非O1/O139株则只具有辅助毒力因子,个别腹泻病例株携带了zot基因。结论:非O1/O139和O139型霍乱弧菌无论在毒力基因种类和质谱特征上都具有显著的差异,广泛应用VITEK-MS微生物快速鉴定技术进行霍乱弧菌血清鉴定有望成为霍乱监测的新助力。Objective:To analysis the regularities of distribution of Vibrio cholera from food risk monitoring samples and evaluate the merits of VITEK-MS rapid analysis methods in identification of Vibrio cholera.Methods:V.cholera strains were isolated from freshwater products and patients.The VITEK-MS and VITEK-2 identification were operated respectively.The serotype was detected by serum agglutination and immunogold.The virulence genes ctxAB was detected by qPCR.All the virulence genes in different types of strains were analyzed by whole genome sequencing.Results:17 of the 23 V.cholera strains were non-O1/O139 serotype.6 strains were detected as O139 V.cholera and ctxAB gene carrier.All the V.cholera strain showed typical mass spectral characteristics based on the fingerprint spectrum.The particular and lacking peaks patterns of O139 V.cholera were representative for Serotype identification.ctxAB,ace and tcp islands genes were harbored in O139 Vibrio cholera,but only auxiliary virulence factor can be detected in non-O1/O139 Vibrio cholera except zot gene.Conclusion:There are many differences between the O139 and non-O1/O139 Vibrio cholera strains in whether virulence gene type or fingerprint spectrum.The thorough application for serotype identification by using microbial mass spectrometry may be a new assistance for cholera surveillance.

关 键 词：霍乱弧菌 微生物质谱(VITEK-MS) 全基因组测序 毒力基因 指纹图谱 

分 类 号：TS201.3[轻工技术与工程—食品科学]