Identification of Penicillium species by MALDI-TOF MS analysis of spores collected by dielectrophoresis
| Autor: | Thelma S. Caine, Michael Alan Reeve, Denise Bachmann |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0303 health sciences
Chromatography biology 030306 microbiology Chemistry Dielectrophoresis biology.organism_classification Mass spectrometry General Biochemistry Genetics and Molecular Biology Spore Agar plate 03 medical and health sciences Matrix-assisted laser desorption/ionization Electrophoresis Penicillium General Agricultural and Biological Sciences Mycelium 030304 developmental biology |
| Zdroj: | Biology Methods and Protocols. 4 |
| ISSN: | 2396-8923 |
| DOI: | 10.1093/biomethods/bpz018 |
| Popis: | In matrix-assisted laser-desorption and ionization mass spectrometry, spectral differences are frequently observed using different growth media on agar plates and/or different growth times in culture, which add undesirable analytical variance. In this article, we explore an approach to the above problem based upon the rationale that, while protein expression in fungal mycelium may well vary under different growth conditions, this might not apply to the same extent in fungal spores. To this end, we have exploited the fact that while mycelium is generally anchored to the fungal-growth substrate, some fungi produce physically-isolated spores which, as such, are amenable to manipulation using dielectrophoresis (the translational motion of charged or uncharged matter caused by polarization effects in a non-uniform electrical field). Such fields can be conveniently generated through the charging of an insulator using the triboelectric effect (the transfer of charge between two objects through friction when they are rubbed together). In this study, polystyrene microbiological inoculating loops were used in combination with nylon-fabric rubbing to harvest fungal spores from five species from within the genus Penicillium, which were grown on agar plates containing two different media over an extended time course. In terms of average Bruker spectral-comparison scores, our method generated higher scores in 80% of cases tested and, in terms of average coefficients of variation, our method generated lower spectral variability in 93% of cases tested. Harvesting of spores using a rapid, inexpensive and simple dielectrophoretic method, therefore, facilitates improved fungal identification for the Penicillium species tested. |
| Databáze: | OpenAIRE |
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