霍乱弧菌基质辅助激光解析电离飞行时间质谱技术谱库的构建  

Establishment of mass spectrometry database of V. cholerae by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

作　　者：张丽荣[1] 黄辉涛 方艳梅[1] 林毅雄 魏泉德[1] 李柏生[2] 

机构地区：[1]珠海市疾病预防控制中心,广东珠海519006 [2]广东省疾病预防控制中心,广东广州511430

出　　处：《中国卫生检验杂志》2018年第2期172-175,共4页Chinese Journal of Health Laboratory Technology

摘　　要：目的利用基质辅助激光解析电离飞行时间质谱(MAIDI-TOF MS)技术构建霍乱弧菌谱库。方法选取56株有代表性的霍乱弧菌,在相同环境下培养,通过生化、分子生物学及血清型鉴定,利用MAIDI-TOF MS对56株霍乱弧菌进行质谱数据采集,建立数据库。另选取10株野生霍乱弧菌及104株非霍乱弧菌用自建数据库进行MAIDI-TOF MS检测及评价。结果自建数据库对10株野生霍乱弧菌鉴定,阳性率为100%,对104株非霍乱弧菌鉴定,均无误判,菌种鉴定结果可靠。56株霍乱菌株的聚类分析发现,MALDI-TOF-MS分型与血清型抗原分类之间无相关性,不同血清型间的质谱数据差异不明显,不能用于区分血清型别。结论本研究建立的霍乱弧菌谱库对霍乱弧菌的鉴定准确性高,适用于实际检测。本研究结果填补了谱库中缺失的霍乱弧菌谱库,建立了霍乱弧菌的MAIDI-TOF MS检测鉴定平台,同时也为其他细菌建立或补充完善谱库提供了很好的方法借鉴。Objective To establish the mass spectrometry database of V. cholerae by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（ MALDI-TOF-MS）. Methods 56 strains of Vibrio cholerae were selected and cultured in the same environment. Through biochemical,molecular biological and serological identification,mass spectrometry data were collected from 56 strains of Vibrio cholerae using MAIDI-TOF MS to establish a database. In addition,10 strains of blind-coded Vibrio cholerae and 104 strains of non-Vibrio cholerae were selected and tested by MAIDI-TOF MS. Results The positive rate of 10 blind-coded V. cholerae strains by the established database was 100%. None of 104 non-V. cholerae strains were identified by the established database. Cluster analysis of 56 cholera strains showed no correlation between MALDI-TOF-MS typing and serotype antigen classification,and there was no significant difference of the spectrum data among different serotypes,therefore,which can not be used to distinguish serotypes. Conclusion The Vibrio cholerae database established in this study is highly accurate for the identification of Vibrio cholerae and is suitable for practical testing. The results of this study have filled the dalabase of Vibrio cholerae deleted in the library and established a platform of MAIDI-TOF MS detection and identification for Vibrio cholerae,and also provided a good reference for other bacteria to establish or supplement the database.

关 键 词：霍乱弧菌 基质辅助激光解析电离飞行时间质谱技术 谱库 

分 类 号：O657.63[理学—分析化学]