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基质辅助激光解吸电离飞行时间质谱在土拉弗朗西斯菌鉴定中的应用

The use of Matrix-assisted laser desorption ionization-time of flight mass spectrometry in the identification of Francisella tularensis.

作者信息

Karatuna Onur, Celebi Bekir, Can Simge, Akyar Isin, Kilic Selcuk

机构信息

Department of Medical Microbiology, Acibadem University School of Medicine, Icerenkoy Mh, Istanbul, Turkey.

出版信息

Bosn J Basic Med Sci. 2016 Jan 15;16(2):132-8. doi: 10.17305/bjbms.2016.894.

Abstract

Francisella tularensis is the cause of the zoonotic disease tularemia and is classified among highly pathogenic bacteria (HPB) due to its low infection dose and potential for airborne transmission. In the case of HBP, there is a pressing need for rapid, accurate and reliable identification. Phenotypic identification of Francisella species is inappropriate for clinical microbiology laboratories because it is time-consuming, hazardous and subject to variable interpretation. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was recently evaluated as a useful tool for the rapid identification of a variety of microorganisms. In this study, we evaluated the use of MALDI-TOF MS for the rapid identification of Francisella tularensis and differentiation of its subspecies. Using national collection of Francisella isolates from the National Tularemia Reference Laboratory (Public Health Institute of Turkey, Ankara), a total of 75 clinical isolates were investigated by species and subspecies-specific polymerase chain reaction (PCR) test and MALDI-TOF MS. All isolates were originally identified as F. tularensis subsp. holarctica due to RD1 subspecies-specific PCR result. For all isolates MALDI-TOF MS provided results in concordance with subspecies-specific PCR analysis. Although PCR-based methods are effective in identifying Francisella species, they are labor-intensive and take longer periods of time to obtain the results when compared with MALDI-TOF MS. MALDI-TOF MS appeared to be a rapid, reliable and cost-effective identification technique for Francisella spp. Shorter analysis time and low cost make this an appealing new option in microbiology laboratories.

摘要

土拉弗朗西斯菌是人畜共患疾病兔热病的病原体,因其感染剂量低且具有空气传播潜力,被归类为高致病性细菌(HPB)。对于高致病性细菌,迫切需要快速、准确和可靠的鉴定方法。土拉弗朗西斯菌的表型鉴定对于临床微生物实验室并不适用,因为它耗时、危险且结果解释存在差异。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)最近被评估为快速鉴定多种微生物的有用工具。在本研究中,我们评估了MALDI-TOF MS在快速鉴定土拉弗朗西斯菌及其亚种方面的应用。利用来自土耳其安卡拉公共卫生研究所国家兔热病参考实验室的土拉弗朗西斯菌分离株国家菌种保藏库,通过种和亚种特异性聚合酶链反应(PCR)试验以及MALDI-TOF MS对总共75株临床分离株进行了研究。由于RD1亚种特异性PCR结果,所有分离株最初都被鉴定为土拉弗朗西斯菌全北区亚种。对于所有分离株,MALDI-TOF MS提供的结果与亚种特异性PCR分析结果一致。尽管基于PCR的方法在鉴定土拉弗朗西斯菌属方面有效,但与MALDI-TOF MS相比,它们劳动强度大且获得结果所需时间更长。MALDI-TOF MS似乎是一种用于土拉弗朗西斯菌属的快速、可靠且经济高效的鉴定技术。更短的分析时间和低成本使其成为微生物实验室中一个有吸引力的新选择。

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