title: Development and optimization of a new MALDI-TOF protocol for identification of the Sporothrix species complex.
abstract: Accurate species identification of the Sporothrix schenckii complex is essential, since identification based only on phenotypic characteristics is often inconclusive due to phenotypic variability within the species. We used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for species identification of 70 environmental and clinical isolates of the Sporothrix complex. A reference database was established for MALDI-TOF MS-based species identification according to minor adjustments in the manufacturer's guidelines. The MALDI-TOF MS clearly distinguished strains of Sporothrix brasiliensis, Sporothrix globosa, Sporothrix mexicana, S. schenckii, Sporothrix luriei and Sporothrix pallida, enabling identification of all isolates at the species level, as confirmed by partial calmodulin gene sequence analyses. The present methodology is simple, reliable, rapid and highly suitable for routine identification in clinical mycology laboratories and culture collections, particularly for updating and reclassifying of deposited Sporothrix isolates.
authors.affiliation: Laboratório de Micologia, Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brazil; CEB-Centre of Biological Engineering, Micoteca da Universidade do Minho, Universidade do Minho, Campus de Gualtar, Braga, Portugal; Centre of Molecular and Environmental Biology (CBMA), Department of Biology, Universidade do Minho, Braga, Portugal. Electronic address: manoel.marques@ini.fiocruz.br.
authors.forename: Manoel Marques Evangelista
authors.initials: MM
authors.lastname: Oliveira
authors.affiliation: CEB-Centre of Biological Engineering, Micoteca da Universidade do Minho, Universidade do Minho, Campus de Gualtar, Braga, Portugal; Programa de Pós-Graduação em Microbiologia Agrícola, Universidade Federal de Lavras, Lavras, MG, Brazil.
authors.forename: Cledir
authors.initials: C
authors.lastname: Santos
authors.affiliation: Centre of Molecular and Environmental Biology (CBMA), Department of Biology, Universidade do Minho, Braga, Portugal.
authors.forename: Paula
authors.initials: P
authors.lastname: Sampaio
authors.affiliation: Department of Environmental and Biological Sciences, University of Messina, Messina, Italy.
authors.forename: Orazio
authors.initials: O
authors.lastname: Romeo
authors.affiliation: Laboratório de Micologia, Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brazil.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Almeida-Paes
authors.affiliation: Centre of Molecular and Environmental Biology (CBMA), Department of Biology, Universidade do Minho, Braga, Portugal.
authors.forename: Célia
authors.initials: C
authors.lastname: Pais
authors.affiliation: CEB-Centre of Biological Engineering, Micoteca da Universidade do Minho, Universidade do Minho, Campus de Gualtar, Braga, Portugal; Programa de Pós-Graduação em Microbiologia Agrícola, Universidade Federal de Lavras, Lavras, MG, Brazil.
authors.forename: Nelson
authors.initials: N
authors.lastname: Lima
authors.affiliation: Laboratório de Micologia, Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brazil.
authors.forename: Rosely Maria
authors.initials: RM
authors.lastname: Zancopé-Oliveira
mesh_terms: D002147:Calmodulin / Q000235:genetics; D019992:Databases as Topic; D005800:Genes, Fungal; D013058:Mass Spectrometry; D010641:Phenotype; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013175:Sporothrix / Q000145:classification* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification
chemical_list: D002147:Calmodulin
keywords: Calmodulin gene; MALDI-TOF MS; Sporothrix complex; Sporotrichosis