Skip to main content

Advertisement

SpringerLink
Log in

Identification of filamentous fungi including dermatophytes using MALDI-TOF mass spectrometry

  • Original Article
  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

Identification of filamentous fungi based on morphological features is the most available approach used in clinical mycology laboratories. However, MALDI-TOF mass spectrometry is currently invaluable for identification of microorganisms because of its rapidity, simplicity, and accuracy. This study aimed to find the optimal way of identifying filamentous fungi using MALDI-TOF MS.

The sample comprised 193 isolates of filamentous fungi. The identification started with morphological assessment. Then isolates were identified using MALDI-TOF MS, both directly from culture and following culture in liquid media with extraction. Subsequently, identification of 20 selected isolates was compared by sequencing of the benA gene, ITS1-5,8-ITS2, and D1-D2 LSU regions.

Based on morphological criteria, 17 genera of fungi were identified. With MALDI-TOF MS performed directly from culture, nine isolates were identified to the genus level and 184 to the species level, with a total of 75 species being noted. With the MALDI-TOF MS extraction method, 190 isolates were identified to the species level, with 43 species being noted. The rates of agreement between identification using morphology and the MALDI-TOF MS direct method were 58.55% at the genus level and 22.24% at the species level. The rates of agreement between identification using morphology and the MALDI-TOF MS extraction method were 84.97% at the genus level and 46.11% at the species level. Using sequencing, 87.5% agreement was found for identification with the MALDI-TOF MS extraction method, as compared with only 43.75% for the direct method.

The results suggest that the optimal approach to identification of filamentous fungi is a combination of morphological features and MALDI-TOF MS using the extraction method.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Almeida F, Rodrigues ML, Coelho C (2019) The still underestimated problem of fungal diseases worldwide. Front Microbiol 10:214. https://doi.org/10.3389/fmicb.2019.00214

    Article  PubMed  PubMed Central  Google Scholar 

  • Bader O (2013) MALDI-TOF-MS-based species identification and typing approaches in medical mycology. Proteomics 13:788–799

    Article  CAS  Google Scholar 

  • Becker PT, de Bel A, Martiny D, Ranque S, Piarroux R, Cassagne C et al (2014) Identification of filamentous fungi isolates by MALDI-TOF mass spectrometry: Clinical evaluation of an extended reference spectra library. Med Mycol 52:826–834

    Article  CAS  Google Scholar 

  • Ciardo DE, Schär G, Altwegg M, Böttger EC, Bosshard PP (2007) Identification of moulds in the diagnostic laboratory - an algorithm implementing molecular and phenotypic methods. Diagn Microbiol Infect Dis 59:49–60

    Article  CAS  Google Scholar 

  • Ciardo DE, Lucke K, Imhof A, Bloemberg GV, Böttger EC (2010) Systematic internal transcribed spacer sequence analysis for identification of clinical mold isolates in diagnostic mycology: a 5-year study. J Clin Microbiol 48:2809–2813

    Article  CAS  Google Scholar 

  • Croxatto A, Prod’hom G, Greub G (2012) Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology. FEMS Microbiol Rev 36:380–407

    Article  CAS  Google Scholar 

  • Demirev PA, Fenselau C (2008) Mass spectrometry for rapid characterization of microorganisms. Annu Rev Anal Chem 1:71–93

    Article  CAS  Google Scholar 

  • Glass NL, Donaldson GC (1995) Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Appl Environ Microbiol 61:1323–1330

    Article  CAS  Google Scholar 

  • Hoenigl M, Salmanton-García J, Walsh TJ, Nucci M, Neoh CF, Jenks JD et al (2020) Global guideline for the diagnosis and management of rare mold infections: an initiative of the ECMM in cooperation with ISHAM and ASM. Lancet Infect Dis. https://doi.org/10.1016/S1473-3099(20)30784-2(inpress)

    Article  PubMed  PubMed Central  Google Scholar 

  • Khot PD, Ko DL, Fredricks DN. Sequencing and analysis of fungal rRNA operons for development of broad-range fungal PCR assays. Appl Environ Microbiol 75:1559–1565

  • Larone DH (2018) Medically important fungi: a guide to identification. ASM Press, Washington DC, p 550

    Google Scholar 

  • Life leading international fungal education (2017) The burden of fungal diseases. New evidence to show the scale of the problem across the globe. http://www.life-worldwide.org/media-centre/article/the-burden-of-fungal-disease-new-evidence-to-show-the-scale-of-the-problem

  • Ranque S, Normand AC, Cassagne C, Murat JB, Bourgeois N, Dalle F et al (2014) MALDI-TOF mass spectrometry identification of filamentous fungi in the clinical laboratory. Mycoses 57:135–140

    Article  CAS  Google Scholar 

  • Rivero-Menendez O, Alastruey-Izquierdo A, Mellado E, Cuenca-Estrella M (2016) Triazole resistance in Aspergillus spp.: a worldwide problem? J Fungi (Basel) 2:21. https://doi.org/10.3390/jof2030021

  • Schuster MG, Cleveland AA, Dubberke ER, Kauffman CA, Avery RK, Husain S et al (2017) Infections in hematopoietic cell transplant recipients: results from the Organ Transplant Infection Project, a multicenter, prospective, cohort study. Open Forum Infect Dis 22:4. https://doi.org/10.1093/ofid/ofx050

    Article  Google Scholar 

  • Theel ES, Hall L, Mandrekar J, Wengenack NL (2011) Dermatophyte identification using matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 49:4067–4071

    Article  CAS  Google Scholar 

  • van Paassen J, Russcher A, In ’t Veld-van Wingerden AW, Verweij PE, Kuijper EJ. (2016) Emerging aspergillosis by azole-resistant Aspergillus fumigatus at an intensive care unit in the Netherlands, 2010 to 2013. Euro Surveill 21(30):2016. https://doi.org/10.2807/1560-7917

    Article  CAS  Google Scholar 

  • Wickes BL, Wiederhold NP (2018) Molecular diagnostics in medical mycology. Nat Commun 3:9. https://doi.org/10.1038/s41467-018-07556-5

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by the grant of the Ministry of Health, the Czech Republic, no. NV 17-31269A. This work was presented as a poster at the 9th Trends in Medical Mycology Congress held on 11–14 October 2019 in Nice, France.

Author information

Authors and Affiliations

  1. Department of Microbiology, University Hospital, Olomouc, Czech Republic

    Petr Hamal & Lucie Svobodova

  2. Department of Microbiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

    Anna Vavrova

  3. Department of Bacteriology and Mycology, Institute of Public Health, Ostrava, Czech Republic

    Jakub Mrazek

Authors
  1. Petr Hamal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anna Vavrova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jakub Mrazek
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lucie Svobodova
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

PH proposed the study design and prepared the final manuscript version. AV identified fungi using MALDI-TOF MS and wrote the first draft of the manuscript. JM identified fungi by the molecular genetic methods and participated in writing of the manuscript. LS performed data analysis and participated in writing of the manuscript.

Corresponding author

Correspondence to Petr Hamal.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hamal, P., Vavrova, A., Mrazek, J. et al. Identification of filamentous fungi including dermatophytes using MALDI-TOF mass spectrometry. Folia Microbiol 67, 55–61 (2022). https://doi.org/10.1007/s12223-021-00917-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12223-021-00917-6

Search

Navigation