Clinical Proteomics Research Center, Chiba University Hospital, 1-8-1 Inohana, Chuo-ku, Chiba City, Chiba, 260-8670, Japan.
Anal Bioanal Chem. 2011 Jun;400(7):1905-11. doi: 10.1007/s00216-011-4877-7. Epub 2011 Mar 26.
In a clinical diagnosis microbiology laboratory, the current method of identifying bacterial isolates is based mainly on phenotypic characteristics, for example growth pattern on different media, colony morphology, Gram stain, and various biochemical reactions. These techniques collectively enable great accuracy in identifying most bacterial isolates, but are costly and time-consuming. In our clinical microbiology laboratory, we prospectively assessed the ability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to identify bacterial strains that were routinely isolated from clinical samples. Bacterial colonies obtained from a total of 468 strains of 92 bacterial species isolated at the Department of Clinical Laboratory at Chiba University were directly placed on target MALDI plates followed by addition of CHCA matrix solution. The plates were then subjected to MALDI-TOF MS measurement and the microorganisms were identified by pattern matching with the libraries in the BioTyper 2.0 software. Identification success at the species and genus levels was 91.7% (429/468) and 97.0% (454/468), respectively. MALDI-TOF MS is a rapid, simple, and high-throughput proteomic technique for identification of a variety of bacterial species. Because colony-to-colony differences and effects of culture duration on the results are minimal, it can be implemented in a conventional laboratory setting. Although for some pathogens, preanalytical processes should be refined, and the current database should be improved to obtain more accurate results, the MALDI-TOF MS based method performs, in general, as well as conventional methods and is a promising technology in clinical laboratories.
在临床诊断微生物学实验室中,目前的细菌分离物鉴定方法主要基于表型特征,例如在不同培养基上的生长模式、菌落形态、革兰氏染色和各种生化反应。这些技术共同实现了对大多数细菌分离物的高度准确性鉴定,但成本高且耗时。在我们的临床微生物学实验室中,我们前瞻性地评估了基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定常规从临床样本中分离的细菌菌株的能力。从千叶大学临床检验科分离的 92 种细菌的 468 株细菌获得的细菌菌落直接置于 MALDI 靶板上,然后加入 CHCA 基质溶液。然后对 MALDI-TOF MS 进行测量,并通过与 BioTyper 2.0 软件中的库进行模式匹配来鉴定微生物。在种和属水平上的鉴定成功率分别为 91.7%(429/468)和 97.0%(454/468)。MALDI-TOF MS 是一种快速、简单、高通量的蛋白质组学技术,可用于鉴定各种细菌。由于菌落之间的差异和培养时间对结果的影响最小,因此可以在常规实验室环境中实施。尽管对于某些病原体,需要细化分析前过程,并改进当前的数据库以获得更准确的结果,但基于 MALDI-TOF MS 的方法总体上与常规方法一样,并且是临床实验室中一种有前途的技术。