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应用基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱法快速鉴定和分型鼠疫耶尔森菌和其他耶尔森氏菌。

Rapid identification and typing of Yersinia pestis and other Yersinia species by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.

机构信息

Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes, UMR CNRS 6236-IRD 198, Faculté de Médecine, IFR48, Université de la Méditerranée, Marseille, France.

出版信息

BMC Microbiol. 2010 Nov 12;10:285. doi: 10.1186/1471-2180-10-285.

DOI:10.1186/1471-2180-10-285
PMID:21073689
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2992509/
Abstract

BACKGROUND

Accurate identification is necessary to discriminate harmless environmental Yersinia species from the food-borne pathogens Yersinia enterocolitica and Yersinia pseudotuberculosis and from the group A bioterrorism plague agent Yersinia pestis. In order to circumvent the limitations of current phenotypic and PCR-based identification methods, we aimed to assess the usefulness of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) protein profiling for accurate and rapid identification of Yersinia species. As a first step, we built a database of 39 different Yersinia strains representing 12 different Yersinia species, including 13 Y. pestis isolates representative of the Antiqua, Medievalis and Orientalis biotypes. The organisms were deposited on the MALDI-TOF plate after appropriate ethanol-based inactivation, and a protein profile was obtained within 6 minutes for each of the Yersinia species.

RESULTS

When compared with a 3,025-profile database, every Yersinia species yielded a unique protein profile and was unambiguously identified. In the second step of analysis, environmental and clinical isolates of Y. pestis (n = 2) and Y. enterocolitica (n = 11) were compared to the database and correctly identified. In particular, Y. pestis was unambiguously identified at the species level, and MALDI-TOF was able to successfully differentiate the three biotypes.

CONCLUSION

These data indicate that MALDI-TOF can be used as a rapid and accurate first-line method for the identification of Yersinia isolates.

摘要

背景

为了将无害的环境耶尔森菌与食源性病原菌小肠结肠炎耶尔森菌和假结核耶尔森菌以及 A 组生物恐怖鼠疫病原体鼠疫耶尔森菌区分开来，准确识别是必要的。为了规避当前表型和基于 PCR 的鉴定方法的局限性，我们旨在评估基质辅助激光解吸/电离飞行时间（MALDI-TOF）蛋白谱分析用于准确快速鉴定耶尔森菌的有用性。作为第一步，我们构建了一个包含 39 种不同耶尔森菌菌株的数据库，代表 12 种不同的耶尔森菌物种，包括 13 种代表 Antiqua、Medievalis 和 Orientalis 生物型的鼠疫耶尔森菌分离株。这些生物在适当的基于乙醇的灭活后被沉积在 MALDI-TOF 板上，并且对于每种耶尔森菌物种都可以在 6 分钟内获得蛋白质谱。

结果

与 3025 个特征谱数据库相比，每种耶尔森菌都产生了独特的蛋白质谱，并能够明确鉴定。在分析的第二步中，环境和临床分离的鼠疫耶尔森菌（n = 2）和小肠结肠炎耶尔森菌（n = 11）与数据库进行了比较并被正确鉴定。特别是，鼠疫耶尔森菌在种水平上被明确鉴定，MALDI-TOF 能够成功区分三种生物型。

结论

这些数据表明，MALDI-TOF 可以用作鉴定耶尔森菌分离株的快速准确的一线方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/27e9/2992509/2cc7e318f469/1471-2180-10-285-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/27e9/2992509/373b48148607/1471-2180-10-285-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/27e9/2992509/2cc7e318f469/1471-2180-10-285-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/27e9/2992509/373b48148607/1471-2180-10-285-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/27e9/2992509/2cc7e318f469/1471-2180-10-285-2.jpg

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