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将Biolog GEN III MicroStation半自动细菌鉴定系统与基质辅助激光解吸电离飞行时间质谱法和16S核糖体RNA基因测序法用于鉴定兽医关注细菌的比较。

Comparison of Biolog GEN III MicroStation semi-automated bacterial identification system with matrix-assisted laser desorption ionization-time of flight mass spectrometry and 16S ribosomal RNA gene sequencing for the identification of bacteria of veterinary interest.

作者信息

Wragg P, Randall L, Whatmore A M

机构信息

Animal Health and Veterinary Laboratories Agency-Penrith Regional Laboratory, Penrith, Cumbria CA11 9RR, United Kingdom.

Department of Bacteriology, Animal Health and Veterinary Laboratories Agency-Weybridge, New Haw, Addlestone, Surrey KT15 3NB, United Kingdom.

出版信息

J Microbiol Methods. 2014 Oct;105:16-21. doi: 10.1016/j.mimet.2014.07.003. Epub 2014 Jul 9.

Abstract

Recent advances in phenotypic and chemotaxonomic methods have improved the ability of systems to resolve bacterial identities at the species level. Key to the effective use of these systems is the ability to draw upon databases which can be augmented with new data gleaned from atypical or novel isolates. In this study we compared the performance of the Biolog GEN III identification system (hereafter, GEN III) with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing in the identification of isolates of veterinary interest. The use of strains that had proven more difficult to identify by routine methods was designed to test the systems' abilities at the extremes of their performance range. Over an 18month period, 100 strains were analysed by all three methods. To highlight the importance of identification to species level, a weighted scoring system was devised to differentiate the capacity to identify at genus and species levels. The overall relative weighted scores were 0.869:0.781:0.769, achieved by 16S rRNA gene sequencing, GEN III and MALDI-TOF MS respectively, when compared to the 'gold standard'. Performance to the genus level was significantly better using 16S rRNA gene sequencing; however, performance to the species level was similar for all three systems.

摘要

表型和化学分类方法的最新进展提高了系统在种水平上分辨细菌身份的能力。有效使用这些系统的关键在于能够利用数据库,这些数据库可以用从非典型或新分离株中收集的新数据进行扩充。在本研究中,我们比较了Biolog GEN III鉴定系统(以下简称GEN III)与基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)以及16S rRNA基因测序在鉴定兽医相关分离株方面的性能。使用常规方法鉴定难度较大的菌株旨在测试这些系统在其性能范围极限时的能力。在18个月的时间里,用这三种方法对100株菌株进行了分析。为了突出种水平鉴定的重要性,设计了一个加权评分系统来区分在属和种水平上的鉴定能力。与“金标准”相比,16S rRNA基因测序、GEN III和MALDI-TOF MS分别获得的总体相对加权分数为0.869:0.781:0.769。使用16S rRNA基因测序在属水平上的性能明显更好;然而,所有三个系统在种水平上的性能相似。

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