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基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)用于常规临床微生物实验室中微生物的快速鉴定。

Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) for rapid identification of micro-organisms in the routine clinical microbiology laboratory.

作者信息

Wattal C, Oberoi J K, Goel N, Raveendran R, Khanna S

机构信息

Sir Ganga Ram Hospital, New Delhi, Delhi, 110060, India.

出版信息

Eur J Clin Microbiol Infect Dis. 2017 May;36(5):807-812. doi: 10.1007/s10096-016-2864-9. Epub 2016 Dec 16.

Abstract

The study evaluates the utility of matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) Vitek MS for identification of microorganisms in the routine clinical microbiology laboratory. From May 2013 to April 2014, microbial isolates recovered from various clinical samples were identified by Vitek MS. In case of failure to identify by Vitek MS, the isolate was identified using the Vitek 2 system (bioMerieux, France) and serotyping wherever applicable or otherwise by nucleic acid-mediated methods. All the moulds were identified by Lactophenol blue mounts, and mycobacterial isolates were identified by molecular identification systems including AccuProbe (bioMerieux, France) or GenoType Mycobacterium CM (Hain Lifescience, Germany). Out of the 12,003 isolates, the Vitek MS gave a good overall ID at the genus and or species level up to 97.7% for bacterial isolates, 92.8% for yeasts and 80% for filamentous fungi. Of the 26 mycobacteria tested, only 42.3% could be identified using the Saramis RUO (Research Use Only) database. VITEK MS could not identify 34 of the 35 yeast isolates identified as C. haemulonii by Vitek 2. Subsequently, 17 of these isolates were identified as Candida auris (not present in the Vitek MS database) by 18S rRNA sequencing. Using these strains, an in-house superspectrum of C. auris was created in the VITEK MS database. Use of MALDI-TOF MS allows a rapid identification of aerobic bacteria and yeasts in clinical practice. However, improved sample extraction protocols and database upgrades with inclusion of locally representative strains is required, especially for moulds.

摘要

本研究评估了基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)Vitek MS在常规临床微生物实验室鉴定微生物中的实用性。2013年5月至2014年4月,从各种临床样本中分离出的微生物菌株通过Vitek MS进行鉴定。若Vitek MS无法鉴定,则使用Vitek 2系统(法国生物梅里埃公司)进行鉴定,并在适用时进行血清分型,否则采用核酸介导的方法进行鉴定。所有霉菌均通过石炭酸品红染色 mounts进行鉴定,分枝杆菌菌株通过包括AccuProbe(法国生物梅里埃公司)或GenoType Mycobacterium CM(德国海因生命科学公司)在内的分子鉴定系统进行鉴定。在12003株分离菌株中,Vitek MS在属和/或种水平上对细菌分离株的总体鉴定成功率高达97.7%,对酵母的鉴定成功率为92.8%,对丝状真菌的鉴定成功率为80%。在测试的26株分枝杆菌中,使用Saramis RUO(仅限研究使用)数据库仅能鉴定出42.3%。VITEK MS无法鉴定Vitek 2鉴定为哈氏假丝酵母的35株酵母分离株中的34株。随后,通过18S rRNA测序,其中17株分离株被鉴定为耳念珠菌(Vitek MS数据库中不存在)。利用这些菌株,在VITEK MS数据库中创建了耳念珠菌的内部超光谱。MALDI-TOF MS的使用可在临床实践中快速鉴定需氧菌和酵母。然而,需要改进样本提取方案并升级数据库,纳入本地代表性菌株,尤其是对于霉菌。

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