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利用质谱法快速鉴定微生物:将内部光谱数据纳入商业数据库可提高性能。

Rapid identification of microorganisms by mass spectrometry: improved performance by incorporation of in-house spectral data into a commercial database.

机构信息

Clinical Proteomics Research Center, Chiba University Hospital, 1-8-1 Inohana, Chiba City, Chiba 260-8670, Japan.

出版信息

Anal Bioanal Chem. 2012 Jun;403(7):1811-22. doi: 10.1007/s00216-011-5656-1. Epub 2011 Dec 27.

Abstract

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is increasingly used as a microbial diagnostic method for species identification of pathogens. However, MALDI-TOF identification of bacteria at the species level remains unsatisfactory, with the major problem being an incomplete database that still needs refinement and expansion. Augmentation of the original MALDI BioTyper 2.0 (Bruker) database by incorporating mass spectra obtained in-house from clinical isolates may increase the identification rate at the species level. We conducted a prospective study to assess whether the augmented database can improve the performance of MALDI-TOF MS for routine identification of species. Cluster analyses revealed distinct differences in MS spectral profiles of clinical isolates obtained in our hospital and those of ATCC strains in the Bruker database. In the first part of the study, which was performed over 3 weeks, 259 bacterial isolates were subjected to analysis by MALDI-TOF MS, and MS spectra of 229 successfully identified isolates (49 species) were incorporated into the original database to give the augmented Bruker-Chiba database. In a second separate analysis, the concordance of identification of 498 clinical isolates of the 49 species with conventional methods was 87.1% (434/498) with the commercial Bruker database and 98.0% (488/498) using the Bruker-Chiba database. These results indicate that refinement of a commercial database can be achieved relatively easy and effectively by incorporating MS spectra of clinical isolates obtained in a clinical laboratory.

摘要

基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)越来越多地被用作微生物诊断方法,用于鉴定病原体的物种。然而,MALDI-TOF 对细菌的物种鉴定仍不尽如人意,主要问题是数据库不完善,仍需要进一步完善和扩展。通过将临床分离株获得的质谱图纳入原始 MALDI BioTyper 2.0(布鲁克)数据库,可以提高物种水平的鉴定率。我们进行了一项前瞻性研究,以评估扩增数据库是否可以提高 MALDI-TOF MS 对常规物种鉴定的性能。聚类分析显示,我们医院获得的临床分离株的 MS 光谱图谱与布鲁克数据库中的 ATCC 菌株存在明显差异。在研究的第一部分(进行了 3 周)中,对 259 株细菌分离株进行 MALDI-TOF MS 分析,并将 229 株成功鉴定的分离株(49 个物种)的 MS 图谱纳入原始数据库,得到扩增的布鲁克-千叶数据库。在第二个独立分析中,使用商业布鲁克数据库对 498 株 49 个物种的临床分离株进行鉴定的一致性为 87.1%(434/498),使用布鲁克-千叶数据库为 98.0%(488/498)。这些结果表明,通过整合临床实验室获得的临床分离株的 MS 图谱,可以相对容易和有效地对商业数据库进行细化。

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