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CryptoType - Public Datasets for MALDI-TOF-MS Based Differentiation of Cryptococcus neoformans/gattii Complexes

Mareike Bernhard1, Navaporn Worasilchai2, Mourine Kangogo3

  • 1Institute for Medical Microbiology, University Medical Center Göttingen, Göttingen, Germany.

Frontiers in Cellular and Infection Microbiology
|May 6, 2021
PubMed

Insights

Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) can accurately identify Cryptococcus species. This study refined sample preparation and created a public database, improving species-level identification for clinical diagnostics.

Area of Science:

  • Medical Mycology
  • Microbiology
  • Mass Spectrometry

Background:

  • Cryptococcus neoformans/gattii species complexes are significant human fungal pathogens, particularly in immunocompromised individuals.
  • Infections range from skin lesions to life-threatening fungal meningitis.
  • Accurate species-level differentiation is crucial due to variations in virulence and antifungal susceptibility.

Purpose of the Study:

  • To re-evaluate sample pre-processing methods for Cryptococcus species identification using MALDI-TOF MS.
  • To establish a publicly available reference library for the MALDI Biotyper system.
  • To improve the accuracy and efficiency of species-level identification in clinical diagnostics.

Main Methods:

  • Four different sample pre-processing protocols were assessed for their impact on peak content.
  • Database entries were created for 13 reference strains.
  • The developed method was validated against 153 clinical isolates previously typed by conventional methods.

Main Results:

  • Standard formic acid extraction with 13 reference entries correctly classified 93.5% (143/153) of clinical isolates.
  • Decapsulating or mechanical disruption protocols did not significantly improve information content.
  • A lowered log score cut-off is suggested due to reproducible identifications in most remaining isolates.

Conclusions:

  • The evaluated workflow enhances the ease and accuracy of species-level identification for Cryptococcus isolates.
  • The created database references are freely available for integration into diagnostic systems.
  • MALDI-TOF MS, with optimized protocols and databases, is a valuable tool for clinical mycology.
Keywords:
Cryptococcus gattii complexCryptococcus neoformans complexMALDI-TOF MScapsuleidentification

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