Donohue Maura J, Smallwood Anthony W, Pfaller Stacy, Rodgers Mark, Shoemaker Jody A
US Environmental Protection Agency, National Exposure Research Laboratory, Cincinnati, OH 45268, USA.
J Microbiol Methods. 2006 Jun;65(3):380-9. doi: 10.1016/j.mimet.2005.08.005. Epub 2005 Sep 19.
This report describes the development of a method to detect the waterborne pathogen Aeromonas using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The genus Aeromonas is one of several medically significant genera that have gained prominence due to their evolving taxonomy and controversial role in human diseases. In this study, MALDI-MS was applied to the characterization of seventeen species of Aeromonas. These seventeen species were represented by thirty-two strains, which included type, reference and clinical isolates. Intact cells from each strain were used to generate a reproducible library of protein mass spectral fingerprints or m/z signatures. Under the test conditions used, peak lists of the mass ions observed in each species revealed that three mass ions were conserved among all the seventeen species tested. These common mass ions having an average m/z of 6301, 12,160 or 12,254, and 13,450, can be potentially used as genus-specific biomarkers to identify Aeromonas in unknown samples. A dendrogram generated using the m/z signatures of all the strains tested indicated that the mass spectral data contained sufficient information to distinguish between genera, species, and strains. There are several advantages of using MALDI-MS based protein mass spectral fingerprinting of whole cells for the identification of microorganisms as well as for their differentiation at the sub-species level: (1) the capability to detect proteins, (2) high throughput, and (3) relatively simple sample preparation techniques. The accuracy and speed with which data can be obtained makes MALDI-MS a powerful tool especially suited for environmental monitoring and detection of biological hazards.
本报告描述了一种使用基质辅助激光解吸/电离质谱(MALDI-MS)检测水传播病原体气单胞菌的方法的开发。气单胞菌属是几个具有医学重要性的属之一,由于其不断演变的分类学以及在人类疾病中存在争议的作用而备受关注。在本研究中,MALDI-MS被应用于对17种气单胞菌的表征。这17个物种由32株菌株代表,其中包括模式菌株、参考菌株和临床分离株。来自每个菌株的完整细胞用于生成可重复的蛋白质质谱指纹图谱或m/z特征库。在所使用的测试条件下,在每个物种中观察到的质量离子峰列表显示,在所有测试的17个物种中,有三个质量离子是保守的。这些平均m/z为6301、12160或12254以及13450的常见质量离子,有可能用作属特异性生物标志物,以识别未知样品中的气单胞菌。使用所有测试菌株的m/z特征生成的树状图表明,质谱数据包含足够的信息来区分属、种和菌株。使用基于MALDI-MS的全细胞蛋白质质谱指纹图谱来鉴定微生物及其在亚种水平上的区分有几个优点:(1)检测蛋白质的能力,(2)高通量,以及(3)相对简单的样品制备技术。能够快速准确地获取数据使得MALDI-MS成为一种特别适用于环境监测和生物危害检测的强大工具。