摘要
目的建立海洋细菌基质辅助激光解吸电离飞行时间质谱鉴定数据库,为海洋细菌引起各类感染的病原快速鉴定提供支持。方法本中心课题组2006-2016年收集国内外6个海域158个采样点海洋细菌530株,分离培养后进行细菌16srDNA扩增测序,应用FlexControl软件收集蛋白谱图,Biotyper软件构建评价数据库。分别用仪器商品化数据库、自建数据库和扩展数据库进行菌种鉴定,比较各数据库的鉴定差异。结果自建本地化海洋细菌鉴定数据库包含32个菌属,82个菌种,共计421株海洋细菌谱图。以16srDNA测序鉴定结果为金标准,验证菌株用商品化数据库鉴定时正确80株,鉴定正确率80.8%,用扩展数据库鉴定时正确97株,鉴定正确率98.0%,差异有统计学意义(P<0.001)。结论自建海洋细菌鉴定数据库菌株的鉴定能力较商品化数据库更强,有助于海洋细菌引起相关感染的诊断和治疗。
Objective To set up marine bacteria identification database by matrix assisted laser desorption ionization time-of-flight mass spectrometry,aiming to provide support for rapid identification of pathogens isolated from marine bacterial infections.Methods Totally 530 marine bacteria were collected by our research team from 6 sea areas and 158 sites from 2006 to 2016.The amplification and sequencing of the 16srDNA of bacteria were performed after isolating and culturing.The FlexControl software was used to detect the protein spectrum and the Biotyper software was used to construct and assess the database.In order to compare if there was difference between the correct identification rates of different databases,the bacterial identification was performed by the commercial,in-house and extended databases,respectively.Results The in-house marine bacteria database included 32 genus,82 species,and 421 strains spectrums in sum.The identification by 16srDNA sequencing was defined as golden standard.Among the validation strains,80 were identified by the commercial database with the accuracy rate of 80.8%,and 97 were identified by extended database(in-house+commercial)with the accuracy rate of 98.0%.The difference was significant(P<0.001).Conclusion Theidentification ability of in-house marine bacteria database is superior to the commercial one and is beneficial to the diagnosis and therapy to the marine bacterial infections.
作者
李艳君
丁毅伟
赵强元
马聪
LI Yanjun;DING Yiwei;ZHAO Qiangyuan;MA Cong(Laboratory Department,the Sixth Medical Center,Chinese PLA General Hospital,Beijing 100048,China)
出处
《解放军医学院学报》
CAS
2020年第7期719-723,共5页
Academic Journal of Chinese PLA Medical School
基金
国家科技重大专项子任务(2018ZX10101003-001-006)
中央军委后勤保障部军队后勤科研重点项目(BHJ13J002)
关键词
基质辅助激光解吸电离飞行时间质谱
海洋细菌
细菌鉴定数据库
病原菌
微生物
matrix assisted laser desorption ionization time-of-flight mass spectrometry
marine bacteria
bacterial identification database
pathogen
microorganism
