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将基质辅助激光解吸电离飞行时间质谱法(MALDI-TOF MS)和MALDI生物分型系统与16S核糖体DNA(rDNA)测序法相比较,用于鉴定从北极海水中分离出的细菌

Evaluation of MALDI-TOF mass spectrometry and MALDI BioTyper in comparison to 16S rDNA sequencing for the identification of bacteria isolated from Arctic sea water.

作者信息

Timperio Anna Maria, Gorrasi Susanna, Zolla Lello, Fenice Massimiliano

机构信息

Dipartimento di Ecologia e Biologia, University of Tuscia, Viterbo, Italy.

Dipartimento di Scienze Agrarie e Forestali, University of Tuscia, Viterbo, Italy.

出版信息

PLoS One. 2017 Jul 24;12(7):e0181860. doi: 10.1371/journal.pone.0181860. eCollection 2017.

Abstract

MALDI-TOF Mass Spectrometry in association with the MALDI BioTyper 3.1 software has been evaluated for the identification and classification of 45 Arctic bacteria isolated from Kandalaksha Bay (White Sea, Russia). The high reliability of this method has been already demonstrated, in clinical microbiology, by a number of studies showing high attribution concordance with other credited analyses. Recently, it has been employed also in other branches of microbiology with controversial performance. The phyloproteomic results reported in this study were validated with those obtained by the "gold standard" 16S rDNA analysis. Concordance between the two methods was 100% at the genus level, while at the species level it was 48%. These percentages appeared to be quite high compared with other studies regarding environmental bacteria. However, the performance of MALDI BioTyper changed in relation to the taxonomical group analyzed, reflecting known identification problems related to certain genera. In our case, attribution concordance for Pseudomonas species was rather low (29%), confirming the problematic taxonomy of this genus, whereas that of strains from other genera was quite high (> 60%). Among the isolates tested in this study, two strains (Exiguobacterium oxidotolerans and Pseudomonas costantinii) were misidentified by MALDI BioTyper due to absence of reference spectra in the database. Accordingly, missing spectra were acquired for the database implementation.

摘要

利用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)结合MALDI BioTyper 3.1软件,对从俄罗斯白海坎达拉克沙湾分离出的45株北极细菌进行了鉴定和分类评估。该方法的高可靠性已在临床微生物学中得到多项研究的证实,这些研究表明其与其他公认分析方法的归因一致性很高。最近,它也被应用于微生物学的其他分支,但其性能存在争议。本研究报告的系统发育蛋白质组学结果与通过“金标准”16S rDNA分析获得的结果进行了验证。两种方法在属水平上的一致性为100%,而在种水平上为48%。与其他关于环境细菌的研究相比,这些百分比似乎相当高。然而,MALDI BioTyper的性能因所分析的分类群而异,这反映了与某些属相关的已知鉴定问题。在我们的案例中,假单胞菌属物种的归因一致性相当低(29%),证实了该属分类的问题,而其他属菌株的归因一致性则相当高(>60%)。在本研究测试的分离株中,由于数据库中没有参考光谱,有两株菌株(耐氧化嗜冷杆菌和康斯坦丁假单胞菌)被MALDI BioTyper误鉴定。因此,为数据库实施获取了缺失的光谱。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/006d/5524297/c9bf3a0fa214/pone.0181860.g001.jpg

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