Ongoing Revolution in Bacteriology: Routine Identification of Bacteria by Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry
Ongoing Revolution in Bacteriology: Routine Identification of Bacteria by Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry
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DOI：
10.1086/600885
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发表时间：
2009-08-15
影响因子：
11.8
通讯作者：
Raoult, Didier
中科院分区：
文献类型：
--
作者：
Seng, Piseth;Drancourt, Michel;Raoult, Didier
背景。基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱法可以准确识别选定的细菌和特定临床情况下的细菌。尚未对其在临床上的常规应用进行评估。我们前瞻性地分析了常规的MALDI-TOF质谱鉴定与传统的细菌表型鉴定，而不考虑门或分离来源。基于16S核糖体RNA和rpoB基因序列的分子鉴定解决了差异。菌落（每个分离物4个点直接沉积在MALDI-TOF板上）使用Autoflex II布鲁克Daltonik质谱仪进行分析。将肽谱与Bruker BioTyper数据库2.0版本进行比较，并记录鉴定分数。测量了识别的延迟和成本。在分析的1660株细菌中，MALDI-TOF质谱法正确率为95.4%；在种水平鉴定的占84.1%，属水平鉴定的占11.3%。在大多数情况下，缺乏鉴定（2.8%的分离株）和错误鉴定（1.7%的分离株）是由于数据库输入不当造成的。准确的MALDI-TOF质谱鉴定与数据库中有10个参考光谱显著相关（P = 0.01）。MALDI-TOF质谱法鉴定1株分离物的平均时间为6分钟，估计成本为目前鉴定方法的22%-32%。MALDI-TOF质谱法是一种经济、准确的常规细菌分离鉴定方法，每个细菌种类有10个参考光谱，鉴定评分>= 1.9 （Brucker系统）。它可能在不久的将来取代革兰氏染色和生化鉴定。
Background. Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry accurately identifies both selected bacteria and bacteria in select clinical situations. It has not been evaluated for routine use in the clinic.Methods. We prospectively analyzed routine MALDI-TOF mass spectrometry identification in parallel with conventional phenotypic identification of bacteria regardless of phylum or source of isolation. Discrepancies were resolved by 16S ribosomal RNA and rpoB gene sequence-based molecular identification. Colonies (4 spots per isolate directly deposited on the MALDI-TOF plate) were analyzed using an Autoflex II Bruker Daltonik mass spectrometer. Peptidic spectra were compared with the Bruker BioTyper database, version 2.0, and the identification score was noted. Delays and costs of identification were measured.Results. Of 1660 bacterial isolates analyzed, 95.4% were correctly identified by MALDI-TOF mass spectrometry; 84.1% were identified at the species level, and 11.3% were identified at the genus level. In most cases, absence of identification (2.8% of isolates) and erroneous identification (1.7% of isolates) were due to improper database entries. Accurate MALDI-TOF mass spectrometry identification was significantly correlated with having 10 reference spectra in the database (P = .01). The mean time required for MALDI-TOF mass spectrometry identification of 1 isolate was 6 minutes for an estimated 22%-32% cost of current methods of identification.Conclusions. MALDI-TOF mass spectrometry is a cost-effective, accurate method for routine identification of bacterial isolates in = 10 reference spectra per bacterial species and a >= 1.9 identification score (Brucker system). It may replace Gram staining and biochemical identification in the near future.