title: Genotyping species of the Sporothrix schenckii complex by PCR-RFLP of calmodulin.
abstract: Sporotrichosis is one of the most common subcutaneous mycosis in Latin America and is caused by 4 pathogenic thermodimorphic fungi in the genus Sporothrix. From both therapeutic and epidemiological perspectives, it is essential to identify the causative agents down to the species level. Traditional parameters may overlap among closely related species, and we propose polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) as an alternative approach. In the present study, the calmodulin gene was amplified and digested with HhaI to yield 5 different electrophoretic patterns representing all medically important Sporothrix species: Sporothrix brasiliensis, Sporothrix schenckii sensu stricto, Sporothrix globosa, and Sporothrix luriei. The PCR-RFLP protocol described here is a simple and inexpensive method and is highly suitable for accurate routine genotyping of relevant Sporothrix species.
authors: Department of Microbiology, Immunology and Parasitology, Cellular Biology Division, Federal University of São Paulo, São Paulo, Brazil; CBS-KNAW Fungal Biodiversity Center, Utrecht, The Netherlands.
authors: Anderson Messias
authors: AM
authors: Rodrigues
authors: CBS-KNAW Fungal Biodiversity Center, Utrecht, The Netherlands.
authors: G Sybren
authors: GS
authors: de Hoog
authors: Department of Microbiology, Immunology and Parasitology, Cellular Biology Division, Federal University of São Paulo, São Paulo, Brazil. Electronic address: zpcamargo@unifesp.br.
authors: Zoilo Pires
authors: ZP
authors: de Camargo
mesh_terms: D002147:Calmodulin / Q000235:genetics*; D006801:Humans; D007843:Latin America; D058889:Molecular Typing / Q000379:methods*; D016533:Mycological Typing Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013175:Sporothrix / Q000145:classification* / Q000235:genetics; D013174:Sporotrichosis / Q000382:microbiology*
chemical_list: D002147:Calmodulin
keywords: Epidemiology; Molecular identification; PCR-RFLP; Sporothrix brasiliensis; Sporothrix schenckii; Sporotrichosis