基质辅助激光解吸电离飞行时间质谱自建库鉴定临床常见致病丝状真菌的研究  

Study on identification of clinical filamentous fungi using MALDI-TOF MS self-built database

作　　者：张璐 赵蓉芬 孙艳霞 Sde Hoog Jos Houbraken 邓淑文 张虹 ZHANG Lu;ZHAO Rongfen;SUN Yanxia;Sde Hoog;Jos Houbraken;DENG Shuwen;ZHANG Hong(Department of Medical Research Center,the People’s Hospital of SND,Suzhou 215004,China;Department of Laboratory Medicine,the People’s Hospital of SND,Suzhou 215004,China;Centre of Expertise in Mycology of Radboud University Medical Centre/Canisius Wilhelmina Hospital,Nijmegen 6500 HB,The Netherlands;Westerdijk Fungal Biodiversity Institute,Utrecht 3508 AD,The Netherlands)

机构地区：[1]苏州高新区人民医院医学研究中心,苏州215004 [2]苏州高新区人民医院检验科,苏州215004 [3]Centre of Expertise in Mycology of Radboud University Medical Centre/Canisius Wilhelmina Hospital,Nijmegen 6500 HB [4]Westerdijk Fungal Biodiversity Institute,Utrecht 3508 AD

出　　处：《广西医科大学学报》2024年第4期500-507,共8页Journal of Guangxi Medical University

基　　金：苏州市科技计划项目(No.SKY2022037);苏州高新区人民医院科学创新基金项目(No.SGY2019B01,No.SGY2021C01,No.SGY2022A01)。

摘　　要：目的:评估基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术鉴定临床常见致病丝状真菌的准确性。方法:采用AUTOF MS1000质谱系统,在原有商业参考库的基础上,增补构建自建数据库,采用125株临床分离菌株对自建库进行验证。结果:自建库新增菌株包括:皮肤癣菌3个属15种菌种,共76株;曲霉属30个菌种,共75株。以已有商业数据库加自建数据库为比对标准,125株临床分离菌株属水平鉴定率100%,种水平鉴定率达80.8%;以分子鉴定结果为金标准,正确鉴定率94.1%。其中,69株皮肤癣菌的种水平鉴定率为79.7%,正确鉴定率92.7%;56株曲霉菌种水平鉴定率为82.1%,正确鉴定率95.6%;错误鉴定6株,集中在须癣毛癣菌复合体(T.mentagrophytes series)菌种趾间毛癣菌/须癣毛癣菌/昆可努发癣菌(T.interdigitale/T.mentagrophytes/T.quinckeanum)交叉识别及Sect.Nigri中黑曲霉(A.niger)与塔宾曲霉(A.tubingensis)交叉识别。结论:通过补充完善自建库可提高MALDI-TOF MS鉴定丝状真菌的准确率;同时,参考数据库及时扩展和更新,以确保MALDI-TOF MS准确鉴定真菌菌种的能力。Objective:To evaluate the accuracy of matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS) in identifying clinical filamentous fungi.Methods:A self-built database was constructed on the basis of the original commercial reference database using AUTOF MS1000 mass spectrometry system,and 125 clinically isolated strains were used to verify it.Results:The self-built database newly added 15species from 3 genera of dermatophytes,totaling 76 strains and 30 species of Aspergillus,totaling 75 strains.The genus-level identification rate of 125 clinically isolated strains was 100% based on existing commercial database and self-built database;the species-level identification rate reached 80.8%.Using the molecular identification as the gold standard,the correct identification rate was 94.1%.Among them,the species-level identification rate of dermatophytes was 79.7%,and the correct identification rate was 92.7%;the species-level identification rate of56 Aspergillus was 82.1%,and the correct identification rate was 95.6%.Six strains were incorrectly identified,concentrated in T.mentagrophytes series whose T.interdigitale,T.mentagrophytes,and T.quinckeanum were cross-identification,and in Sect.Nigri whose A.niger and A.tubingensis were cross-identification.Conclusion:The accuracy of MALDI-TOF MS identification of filamentous fungi can be improved by supplementing and perfecting the self-built database.Meanwhile,the reference database should be expanded and updated in time to ensure the ability of MALDI-TOF MS to accurately identify fungal strains.

关 键 词：基质辅助激光解吸电离飞行时间质谱 自建库 皮肤癣菌 曲霉 

分 类 号：R519[医药卫生—内科学]