基于MALDI-TOF MS的弧菌识别能力分析  

Analysis of the ability of MALDI-TOF MS to identify Vibrio

作　　者：张慧芳 李莉[2] 麻丽丹 张建中 肖迪 

机构地区：[1]中国疾病预防控制中心传染病预防控制所,中国疾病预防控制中心传染病预防控制国家重点实验室和感染性疾病诊治协同创新中心,北京102206 [2]辽宁出入境检验检疫局,辽宁大连116001 [3]丹东出入境检验检疫局,辽宁丹东118000

出　　处：《中国病原生物学杂志》2016年第6期522-529,534,共9页Journal of Pathogen Biology

基　　金：国家质检总局科研计划项目(No.2012IK170);传染病预防控制国家重点实验室自主项目(No.2014SKLID102);中国疾病预防控制中心传染病预防控制所科研课题

摘　　要：目的确定基质辅助激光解析飞行时间质谱(MALDI-TOF MS)-Biotyper系统对弧菌属细菌的识别能力。方法采用Bruker Microflex LT系统,通过构建30株弧菌的肽质量参考谱库、完善Biotyper数据库;采用77株弧菌对数据库的识别能力进行评价;采用主谱图预测聚类分析揭示其对弧菌属、种水平的区分能力及肽质量谱与菌株毒力的相关性。结果 107株弧菌通过MALDI-TOF MS-Biotyper原数据库系统鉴定,92.5%(99株)为属水平识别,55.1%(59株)为种水平识别。用于外部验证的77株菌搜索Biotyper原数据库和补充完善后的数据库,种水平识别率从53.2%提高至96.1%,提高42.9%;属水平识别率从92.2%提高至100%,提高7.8%;聚类分析显示弧菌属中麦氏弧菌与溶藻弧菌、弗尼斯弧菌肽质量谱特异性差,是造成识别错误的主要因素;聚类信息显示,肽质量谱与霍乱弧菌的产毒性不相关,与副溶血性弧菌毒力不相关。结论 MALDI-TOF MS-Biotyper系统可用于弧菌属、种水平的快速识别。数据库的完善程度是决定种水平识别能力的关键。Objective To determine if a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MS MALDI-TOF）-Biotyper system is able to identify Vibrio species. Methods Reference mass spectra for peptides （30 strains of Vibrio） were obtained from the Bruker Microflex LT system and supplemented with profiles in the Biotyper database. The ability of the database to identify Vibrio species was evaluated with 77 strains of Vibrio. Main spectra pro- jection and clustering analysis were used to distinguish strains at the genus and species level, and the correlation between the mass spectra for peptides and the virulence of the strains was analyzed. Results One hundred and seven strains of Vibrio were identified by the MALDI-TOF MS-Biotyper database system; 99 strains （92.5 %） were accurately identified at the genus level and 59 strains （55.1%） were accurately identified at the species level. For external validation, 77 bacterial strains in the original reference database and the supplemented database were compared. The supplemented database improved identification 7.8% at the genus level （from 92.2% to 100%） and 42.9% at the species level （from 53.2% to 96.1 %）. Clustering analysis indicated differences in the specificity of mass spectra for peptides of Vibrio metschnikovii, V. alginolyticus, and V. furnissii according to Biotyper, and this was the main cause of identification error. Clustering information also indicated that the mass spectra for peptides were not related to the virulence of V. cholera or V. parahernolyticus. Conclusion The MALDI-TOF MS-Biotyper system can be used to rapidly identify Vibrio at the genus and species level. Supplementing the database is the key to determining the system＇s ability to identify different species

关 键 词：基质辅助激光解析飞行时间质谱 弧菌 识别能力 特异性 

分 类 号：R378.3[医药卫生—病原生物学]