The development of a matrix-assisted laser desorption/ionization mass spectrometry-based method for the protein fingerprinting and identification of Aeromonas species using whole cells

被引:52
作者
Donohue, Maura J.
Smallwood, Anthony W.
Pfaller, Stacy
Rodgers, Mark
Shoemaker, Jody A.
机构
[1] US EPA, Natl Exposure Res Lab, Cincinnati, OH 45268 USA
[2] US EPA, Natl Exposure Res Lab, Natl Council Aging, Cincinnati, OH 45268 USA
[3] US EPA, Natl Risk Management Res Lab, Cincinnati, OH 45268 USA
关键词
Aeromonas; MALDI-MS; microorganism; whole cell analysis;
D O I
10.1016/j.mimet.2005.08.005
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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-NIS 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. Published by Elsevier B.V.
引用
收藏
页码:380 / 389
页数:10
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