Abstract
MALDI-TOF MS has become the standard method for routine identification of most microbial organisms in clinical laboratories and has largely replaced biochemical assays. Classification relies on extensive well curated databases, ideally covering the full spectrum of microorganisms encountered in the specimens at hands. The protocols for harvesting cells and procuring material suitable for downstream MALDI-TOF MS analyses vary in specific details between the different groups of organisms, e.g., gram-positive or -negative bacteria, mycobacteria, or fungi. With respect to fungi, methods further vary between yeasts and moulds; and even among different mould genera if they do not lyse in a similar fashion. Purification of microbial materials from clinical specimen allows the direct identification of bacteria; however this is not yet fully adapted to fungi. In this chapter, I look into the differences between the underlying methods for yeast and moulds, and for production of samples suitable for MALDI-TOF MS species identification from cultures and different clinical materials.
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Acknowledgements
The technique itself and the basic protocols outlined here have been developed over the years by many different people. I apologize to those that go unmentioned. Specifically I would like to thank my coworkers Andreas Zautner and Michael Weig, as well as Thomas Maier (Bruker Daltonics) and Marcel Erhard (Anagnostec and bioMérieux) and their teams for many discussions on the topic, and the repeated personal teaching and problem-solving efforts made. The figures were created and kindly contributed by Maram Bader.
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Bader, O. (2017). Fungal Species Identification by MALDI-ToF Mass Spectrometry. In: Lion, T. (eds) Human Fungal Pathogen Identification. Methods in Molecular Biology, vol 1508. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6515-1_19
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DOI: https://doi.org/10.1007/978-1-4939-6515-1_19
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