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开发一个用于通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)快速准确地常规鉴定无色杆菌属菌种的数据库。

Development of a database for the rapid and accurate routine identification of Achromobacter species by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS).

作者信息

Garrigos T, Neuwirth C, Chapuis A, Bador J, Amoureux L

机构信息

Department of Bacteriology, University Hospital of Dijon, BP 37013, 21070, Dijon Cedex, France.

Department of Bacteriology, University Hospital of Dijon, BP 37013, 21070, Dijon Cedex, France; UMR/CNRS 6249 Chrono-environnement, University of Bourgogne- Franche-Comté, Besançon, France.

出版信息

Clin Microbiol Infect. 2021 Jan;27(1):126.e1-126.e5. doi: 10.1016/j.cmi.2020.03.031. Epub 2020 Apr 10.

DOI:10.1016/j.cmi.2020.03.031
PMID:32283265
Abstract

OBJECTIVES

Achromobacter spp. are emerging pathogens in respiratory samples from cystic fibrosis patients. The current reference methods (nrdA-sequencing or multilocus sequence typing) can identify 18 species which are often misidentified by conventional techniques as A. xylosoxidans. A few studies have suggested that matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) provides accurate identification of the genus but not of species. The aims of this study were (a) to generate a database for MALDI-TOF/MS Bruker including the 18 species, (b) to evaluate the suitability of the database for routine laboratory identification, and (c) to compare its performance with that of the currently available Bruker default database.

METHODS

A total of 205 isolates belonging to the 18 species identified by nrdA sequencing were used to build a local database. Main spectra profiles (MSPs) were created according to Bruker's recommendations for each isolate with the Biotyper software. Performance of the default Bruker database and ours for routine use were compared by testing 167 strains (including 38 isolates used from MSP creation) belonging to the 18 species identified by nrdA sequencing directly from colonies cultivated on various media.

RESULTS

Our new database accurately identified 99.4% (166/167) of the isolates from the 18 species (score ≥2.0) versus only 50.9% (85/167) with the Bruker database. In the Bruker database 17.3% of the isolates (29/167) were incorrectly identified as another species despite a score of ≥2.0.

CONCLUSIONS

The use of MALDI-TOF/MS in combination with a database developed with samples from 18 Achromobacter species provides rapid and accurate identification. This tool could be used to help future clinical studies.

摘要

目的

无色杆菌属是囊性纤维化患者呼吸道样本中新兴的病原体。当前的参考方法(nrdA测序或多位点序列分型)可鉴定出18个物种,这些物种常被传统技术误鉴定为木糖氧化无色杆菌。一些研究表明,基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF/MS)可准确鉴定该属,但无法鉴定物种。本研究的目的是:(a)为布鲁克MALDI-TOF/MS生成一个包含这18个物种的数据库;(b)评估该数据库用于常规实验室鉴定的适用性;(c)将其性能与当前可用的布鲁克默认数据库进行比较。

方法

共使用205株通过nrdA测序鉴定的属于这18个物种的分离株构建本地数据库。根据布鲁克的建议,使用Biotyper软件为每个分离株创建主要光谱图谱(MSP)。通过直接测试来自在各种培养基上培养的菌落的167株菌株(包括38株用于创建MSP的分离株),比较布鲁克默认数据库和我们的常规使用数据库的性能,这些菌株属于通过nrdA测序鉴定的18个物种。

结果

我们的新数据库准确鉴定了18个物种中99.4%(166/167)的分离株(分数≥2.0),而使用布鲁克数据库时仅为50.9%(85/167)。在布鲁克数据库中,17.3%的分离株(29/167)尽管分数≥2.0,但仍被错误鉴定为另一个物种。

结论

将MALDI-TOF/MS与使用来自18种无色杆菌物种的样本开发的数据库结合使用,可实现快速准确的鉴定。该工具可用于辅助未来的临床研究。

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