Analytical and Bioanalytical Chemistry

, Volume 408, Issue 27, pp 7565–7581 | Cite as

Identification of Stachybotrys spp. by MALDI-TOF mass spectrometry

  • Sebastian Ulrich
  • Barbara Biermaier
  • Oliver Bader
  • Georg Wolf
  • Reinhard K. Straubinger
  • Andrea Didier
  • Brigitte Sperner
  • Karin Schwaiger
  • Manfred Gareis
  • Christoph Gottschalk
Paper in Forefront
First Online:
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Revised:
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Abstract

Stachybotrys (S.) spp. are omnipresent cellulolytic molds. Some species are highly toxic owing to their ability to synthesize various secondary metabolites such as macrocyclic trichothecenes or hemolysins. The reliable identification of Stachybotrys at species level is currently limited to genome-based identification. This study aimed to establish a fast and reliable MALDI-TOF MS identification method by optimizing the pre-analytical steps for protein extraction for subsequent generation of high-quality fingerprint mass spectra. Eight reference strains of the American Type Culture Collection and the Technical University of Denmark were cultivated in triplicate (biological repetitions) for 2 days in malt extract broth. The mycelia (1.5 ml) were first washed with 75 % ethanol and an additional washing step with dimethyl sulfoxide (10 %) was added to remove unspecific low weight masses. Furthermore, mycelia were broken with roughened glass beads in formic acid (70 %) and acetonitrile. The method was successfully applied to a total of 45 isolates of Stachybotrys originating from three different habitats (indoor, feed, and food samples; n = 15 each): Twenty-seven isolates of S. chartarum and 18 isolates of S. chlorohalonata could be identified by MALDI-TOF MS. The data obtained exactly matched those obtained by genome-based identification. The mean score values for S. chartarum ranged from 2.509 to 2.739 and from 2.148 to 2.622 for S. chlorohalonata with a very good reproducibility: the relative standard deviations were between 0.3 % and 6.8 %. Thus, MALDI-TOF MS proved to be a fast and reliable alternative to identification of Stachybotrys spp. by nucleotide amplification and sequencing.

Keywords

Stachybotrys spp MALDI-TOF MS Mass spectrometry Filamentous fungi 
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Notes

Acknowledgements

We are especially grateful to the Brigitte and Wolfram Gedek foundation for the financial support of this research work. Part of the work was supported by the German Academic Exchange Service (DAAD), project number 56269877.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Sebastian Ulrich
    • 1
  • Barbara Biermaier
    • 1
  • Oliver Bader
    • 2
  • Georg Wolf
    • 3
  • Reinhard K. Straubinger
    • 3
  • Andrea Didier
    • 4
  • Brigitte Sperner
    • 1
  • Karin Schwaiger
    • 1
  • Manfred Gareis
    • 1
  • Christoph Gottschalk
    • 1
  1. 1.Chair of Food Safety, Department of Veterinary SciencesLMU MunichOberschleissheimGermany
  2. 2.Institute for Medical MicrobiologyUniversity Medical Center GoettingenGoettingenGermany
  3. 3.Chair of Microbiology and Mycology, Department of Veterinary SciencesLMU MunichMunichGermany
  4. 4.Chair for Hygiene and Technology of Milk, Department of Veterinary SciencesLMU MunichOberschleissheimGermany

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