Vol2-2Art5 Clinical use of MALDI-ToF MS for Sporothrix ID

Mariana Cruzetta de Carvalho¹, Germana Davila dos Santos², Marisol Dominguez Muro^3,7, Bram Spruijtenburg^4,5,6, Carolina Melchior do Prado^5,6,7, Norma B. Fernandez⁹, Izabella Castilhos Ribeiro dos Santos Weiss¹⁰, Eelco F.J. Meijer^4,5,6, Flávio de Queiroz-Telles¹¹, Vânia Aparecida Vicente^2,7,8#, Regielly Caroline Raimundo Cognialli^3,7#

¹Postgraduate Residency Program in Oncology and Hematology, Hospital de Clínicas, Federal University of Paraná, Curitiba, Brazil; ²Postgraduate Program in Engineering Bioprocess and Biotechnology, Federal University of Paraná, Curitiba, Brazil;³Hospital de Clínicas, Federal University of Paraná, Curitiba, Brazil; ⁴Department of Medical Microbiology, Radboudumc, Nijmegen, The Netherlands; ⁵Radboudumc-CWZ Center of Expertise for Mycology, Nijmegen, The Netherlands; ⁶Department of Medical Microbiology and Immunology, Canisius-Wilhelmina Hospital (CWZ) / Dicoon, Nijmegen, The Netherlands; ⁷Postgraduate Program in Microbiology, Parasitology and Pathology, Federal University of Paraná, Curitiba, Brazil; ⁸Department of Basic Pathology, Federal University of Paraná, Curitiba, Brazil; ⁹Hospital de Clínicas José de San Martin, UBA, Ciudad Autónoma de Buenos Aires, Argentina; ¹⁰Department of Clinical Analysis, Federal University of Paraná, Curitiba, Brazil; ¹¹Department of Public Health, Federal University of Paraná, Curitiba, Brazil

Abstract

Sporotrichosis, a neglected mycosis caused by thermodimorphic Sporothrix species, is of growing concern due to the virulent S. brasiliensis, responsible for a large Brazilian outbreak with regional and international spread. Molecular tools remain the identification gold standard, Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-ToF MS) offers a rapid proteomic alternative. Therefore, in this study we developed and validated an in-house MALDI-TOF MS database for S. brasiliensis and S. schenckii using the Bruker Biotyper MBT Smart system. Spectra from mycelial and yeast phases of 14 clinical strains (11 S. brasiliensis, 3 S. schenckii) were used for database construction; 40 additional strains were tested for validation. Mycelial-phase spectra showed interspecies overlap, whereas yeast-phase profiles exhibited discriminatory peaks (6,000–8,000 Da). ROC analysis indicated day 5 as the earliest reliable timepoint (58 % identifications, score ≥2.15) and optimal performance at day 12 (85 % identifications, score ≥1.91). MALDI-ToF MS is a promising complementary tool for Sporothrix identification in endemic areas. Broader multicenter datasets incorporating diverse species and geographic variants are needed to enhance diagnostic accuracy. 

#Correspondence:
regielly.cognialli@gmail.com
vaniava63@gmail.com 

Article info:
Received 27 September 2025 
Revised 20 October 2025 
Accepted 25 October 2025
Appeared online 30 October 2025
One Health Mycology 2(2): 142-151, 2025
doi.org/10.63049/OHM.25.22.5 

Cite this article:
de Carvalho MC, dos Santos GD , Dominguez Muro M, Spruijtenburg B, Melchior do Prado C, Fernandez NB, Ribeiro dos Santos Weiss IC, Meijer EFJ, de Queiroz-Telles F, Vicente VA, Cognialli RCR 2025. Potential clinical application of MALDI-ToF MS using the Bruker Biotyper for the identification of Sporothrix brasiliensis and Sporothrix schenckii. One Health Mycology 2(2), 142-151
This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/ licenses/by-nc-nd/4.0/)

Keywords:
sporotrichosis, mass spectrometry, diagnostics, identification

Article highlights:

• An in-house MALDI-ToF MS database was established using both mycelial and yeast-phase spectra of Sporothrix brasiliensis and S. schenckii.
• Validation with 40 strains showed temporal differences across days of culture, and application of the optimized protocol enabled correct species-level identification in 85% of Sporothrix strains. 
• MALDI-ToF MS constitutes a promising complementary diagnostic approach for identification of Sporothrix species in endemic regions.