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基于 MALDI-TOF 质谱技术的大肠杆菌血清型鉴定方法评估

Evaluation of MALDI-TOF mass spectroscopy methods for determination of Escherichia coli pathotypes.

机构信息

Enteric Diseases Program, Bacteriology and Enteric Diseases Program, National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB R3E 3R2 Canada.

出版信息

J Microbiol Methods. 2013 Sep;94(3):180-91. doi: 10.1016/j.mimet.2013.06.020. Epub 2013 Jun 28.

Abstract

It is rapidly becoming apparent that many E. coli pathotypes cause a considerable burden of human disease. Surveillance of these organisms is difficult because there are few or no simple, rapid methods for detecting and differentiating the different pathotypes. MALDI-TOF mass spectroscopy has recently been rapidly and enthusiastically adopted by many clinical laboratories as a diagnostic method because of its high throughput, relatively low cost, and adaptability to the laboratory workflow. To determine whether the method could be adapted for E. coli pathotype differentiation the Bruker Biotyper methodology and a second methodology adapted from the scientific literature were tested on isolates representing eight distinct pathotypes and two other groups of E. coli. A total of 136 isolates was used for this study. Results confirmed that the Bruker Biotyper methodology that included extraction of proteins from bacterial cells was capable of identifying E. coli isolates from all pathotypes to the species level and, furthermore, that the Bruker extraction and MALDI-TOF MS with the evaluation criteria developed in this work was effective for differentiating most pathotypes.

摘要

显然,许多大肠杆菌血清型会对人类健康造成严重负担。由于缺乏简单、快速的方法来检测和区分不同的血清型,因此对这些病原体的监测具有一定难度。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)最近已被许多临床实验室迅速采用,作为一种诊断方法,因为其具有高通量、相对低成本和适应实验室工作流程的特点。为了确定该方法是否可用于大肠杆菌血清型的区分,本研究测试了 Bruker Biotyper 方法和从文献中改编的第二种方法,分别对 8 种不同血清型和另外 2 组大肠杆菌的分离株进行了检测。该研究共使用了 136 株分离株。结果证实,包括从细菌细胞中提取蛋白质的 Bruker Biotyper 方法能够将所有血清型的大肠杆菌分离株鉴定到种水平,而且,Bruker 提取法和 MALDI-TOF MS 结合本研究中制定的评估标准,可有效区分大多数血清型。

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