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基质辅助激光解吸电离飞行时间质谱法和基质辅助生物型鉴定受污染辣根根际土壤中培养的联苯代谢细菌。

Matrix-assisted laser desorption ionization (MALDI)-time of flight mass spectrometry- and MALDI biotyper-based identification of cultured biphenyl-metabolizing bacteria from contaminated horseradish rhizosphere soil.

机构信息

Institute of Chemical Technology Prague, Faculty of Food and Biochemical Technology, Department of Biochemistry and Microbiology, Technicka 3, 166 28 Prague, Czech Republic.

出版信息

Appl Environ Microbiol. 2011 Oct;77(19):6858-66. doi: 10.1128/AEM.05465-11. Epub 2011 Aug 5.

Abstract

Bacteria that are able to utilize biphenyl as a sole source of carbon were extracted and isolated from polychlorinated biphenyl (PCB)-contaminated soil vegetated by horseradish. Isolates were identified using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The usage of MALDI Biotyper for the classification of isolates was evaluated and compared to 16S rRNA gene sequence analysis. A wide spectrum of bacteria was isolated, with Arthrobacter, Serratia, Rhodococcus, and Rhizobium being predominant. Arthrobacter isolates also represented the most diverse group. The use of MALDI Biotyper in many cases permitted the identification at the level of species, which was not achieved by 16S rRNA gene sequence analyses. However, some isolates had to be identified by 16S rRNA gene analyses if MALDI Biotyper-based identification was at the level of probable or not reliable identification, usually due to a lack of reference spectra included in the database. Overall, this study shows the possibility of using MALDI-TOF MS and MALDI Biotyper for the fast and relatively nonlaborious identification/classification of soil isolates. At the same time, it demonstrates the dominant role of employing 16S rRNA gene analyses for the identification of recently isolated strains that can later fill the gaps in the protein-based identification databases.

摘要

从辣根污染的多氯联苯 (PCB) 污染土壤中提取并分离出能够将联苯作为唯一碳源利用的细菌。使用基质辅助激光解吸电离飞行时间质谱 (MALDI-TOF MS) 鉴定分离物。评估并比较了 MALDI Biotyper 对分离物的分类与 16S rRNA 基因序列分析的用途。分离出了广泛的细菌,其中以节杆菌属、沙雷氏菌属、红球菌属和根瘤菌属为主。节杆菌属分离物也代表了最多样化的群体。在许多情况下,MALDI Biotyper 的使用可以在种的水平上进行鉴定,而 16S rRNA 基因序列分析无法达到这一水平。然而,如果 MALDI Biotyper 基于鉴定的水平为可能或不可靠鉴定,则必须通过 16S rRNA 基因分析进行鉴定,通常是由于数据库中缺少参考光谱。总的来说,这项研究表明,使用 MALDI-TOF MS 和 MALDI Biotyper 可以快速且相对不费力地鉴定/分类土壤分离物。同时,它证明了采用 16S rRNA 基因分析对于鉴定新分离的菌株的主导作用,这些菌株可以填补基于蛋白质的鉴定数据库中的空白。

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