Trichosporon asahii is an opportunistic fungus considered the leading etiologic agent of trichospornosis, a disease that causes great morbidity/mortality among affected patients. The identification of the etiologic agent is generally obtained through physiological and morphological studies. Molecular investigations, such as species-specific primers (PCR), have recently been developed with the aim of applying a more simple, specific, and faster technology for mycological diagnosis. The genetic material amplification technique using ad-random primers (RAPD: random amplified polymorphic DNA) is an epidemiological tool which enables research on infection by and transmission of suspected agents. In this study, the amplified polymorphic DNA technique was used to determine the intraspecific diversity of 10 Trichosporon asahii strains. Primers OPAO-15 and 1821 were used and these allowed association to 5 and 3 electrophoretic patterns, respectively. The T. asahii molecular identification, which had been previously analyzed by conventional methods, was performed by means of primers TAAF and pITS4. Our results support the use of these techniques for clonality studies of the strains of this fungus as well as for the fast and specific identification of its members in clinical cases.