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利用基质辅助激光解吸电离飞行时间质谱法鉴定动物巴斯德氏菌科细菌

Identification of animal Pasteurellaceae by MALDI-TOF mass spectrometry.

作者信息

Frey Joachim, Kuhnert Peter

机构信息

Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Bern, Bern, Switzerland,

出版信息

Methods Mol Biol. 2015;1247:235-43. doi: 10.1007/978-1-4939-2004-4_18.

Abstract

Species of the family Pasteurellaceae play an important role as primary or opportunistic, predominantly respiratory, pathogens in domestic and wild animals. Some of them cause severe disease with high economic losses in commercial animal husbandry. Hence, rapid and accurate differentiation of Pasteurellaceae is important and signifies a particular challenge to diagnostic laboratories. Identification and differentiation of Pasteurellaceae is mostly done using phenotypic tests or genetic identification based on sequence similarity of housekeeping genes, such as the rrs gene encoding the 16S ribosomal RNA (16S rRNA). Both approaches are time consuming, laborious, and costly, therefore often delaying the final diagnosis of disease or epidemics. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry represents an alternative rapid and reliable method for the differentiation of most members of the family Pasteurellaceae. It is able to differentiate within a few minutes the currently known 18 genera and most of the over 60 species and subspecies of Pasteurellaceae including many members encountered in veterinary diagnostic laboratories. A few closely related species and subspecies that cannot be discriminated by MALDI-TOF are easily identified further by complementary simple tests, such as hemolysis done simultaneously or routinely during pathogen isolation.

摘要

巴斯德氏菌科的菌种作为家畜和野生动物中的原发性或机会性主要呼吸道病原体发挥着重要作用。其中一些会引发严重疾病,给商业畜牧业造成巨大经济损失。因此,快速准确地区分巴斯德氏菌科很重要,这对诊断实验室来说是一项特殊挑战。巴斯德氏菌科的鉴定和区分大多通过表型测试或基于管家基因序列相似性的基因鉴定来完成,比如编码16S核糖体RNA(16S rRNA)的rrs基因。这两种方法都耗时、费力且成本高,因此常常会延误疾病或疫情的最终诊断。基质辅助激光解吸/电离飞行时间(MALDI-TOF)质谱分析法是区分巴斯德氏菌科大多数成员的另一种快速可靠的方法。它能够在几分钟内区分出目前已知的18个属以及巴斯德氏菌科60多个种和亚种中的大多数,包括兽医诊断实验室中遇到的许多成员。一些无法通过MALDI-TOF区分的亲缘关系较近的种和亚种,通过补充一些简单测试很容易进一步鉴定出来,比如在病原体分离过程中同时或常规进行的溶血试验。

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