121 ECCO XXXI MEETING | ABSTRACTS BOOK P59 MALDI-TOF ICMS: A PROTEOMIC METHOD FOR IDENTIFICATION OF CLINICAL SPOROTHRIX COMPLEX ISOLATES M. M. E. Oliveira 1,2,3 , C. Santos 2 , P. Sampaio 3 , C. Pais 3 , N. Lima 2 and R. M. Zancopé-Oliveira 1 1 Laboratório de Micologia, Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brazil 2 IBB/Centre of Biological Engineering, Micoteca da Universidade do Minho, Braga, Portugal 3 Molecular and Environmental Biology Centre (CBMA), Department of Biology, Universidade do Minho, Braga, Portugal e-mail: rosely.zancope@ipec.fiocruz.br Sporotrichosis is a subcutaneous mycosis of worldwide distribution. However, Latin America, South Africa, India and Japan are areas of high endemicity. Recently, a combination of phenotypic and genotypic features suggested that Sporothrix schenckii should not be considered as a single taxon causing sporotrichosis as 3 new species, S. brasiliensis, S. globosa and S. mexicana have recently been described. Sporothrix mex- icana was related with environmental samples and apparently restricted to Mexico. However, our Research Group has recently described the first case of human sporotri- chosis caused by S. mexicana in Portugal [1]. An identification key for the Sporothrix complex species has now been proposed which includes macro- and micro-morphology and auxonogram analyses using raffinose and sucrose as carbon sources. Neverthe- less, identification based on this methodology could be ambiguous due to phenotypic variability within these species [2]. In addition, conclusive species identification is reached only after partial calmodulin gene (CAL) sequence analysis. In order to show the potential of the Matrix-Assisted Laser Desorption/Ionisation Time-Of-Flight In- tact Cell Mass Spectrometry (MALDI-TOF ICMS) technique on the identification of Sporothrix complex species the aim of this study was to optimise the MALDI-TOF ICMS methodology for the 4 available Sporothrix isolates related with human sporo- trichosis. For that proposal the type strain S. brasiliensis IPEC16490 (CBS 120339) and the reference strains S. globosa IPEC27135, S. schenckii IPEC27722 and S. mexicana MUM11.02 were used. Also were compared this isolates in two morphologic phases. The analysis demonstrated that optimal spectra and statistical clustering were obtained when the microbial cells were analysed on the yeast phase. The present methodology is simple, reliable, and rapid making it an ideal routine identification system for clinical mycology laboratories and culture collections. Acknowledgements: M.M.E. Oliveira was supported by a grant from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Brazil. We would like to thank Marília Maciel for her contribution on MALDI-TOF ICMS analysis. References: [1] Dias, N., Oliveira, M.M.E., Portela, M., Santos, C., Zancopé-Oliveira, R.M., Lima, N. (2011) Human sporotri- chosis caused by Sporothrix mexicana in a Portuguese patient. Emerging Infectious Diseases, 17:1975-1976. [2] Oliveira, M.M.E., Almeida-Paes, R., Muniz, M.M., Gutierrez-Galhardo, M.C., Zancope-Oliveira, R.M. (2011) Phenotypic and molecular identification of Sporothrix isolates from an epidemic area of sporotrichosis in Brazil. Mycopathologia,169:359–363.