FIGURE 2: A 7% polyacrylamide gel stained with 2% silver nitrate showing the nested-PCR (In) amplification using the primer pair SPOR3 (forward) (5′-ATCTCTTGGCTCTGGCATCG-3′) and R4 (reverse) (5′-GGGAGAACATGCGTTCGGTA-3′), with 1 ?#61549;L of the amplicon of the first PCR (“Out”), diluted to 1/10, as the DNA template. The fragment amplified was approximately 260 bp, as observed via computational tool analysis. For the PCR specificity test, strains of different fungal species were analyzed. Lane 1: molecular weight standard, 100 bp ladder; lane 2: DNA extracted from an international standard sample of Sporothrix schenkii ATCC MF765977 as a positive control for the reaction; lanes 3-4: DNA extracted from biopsies of patients presenting with sporotrichosis confirmed by culture, showing bands of approximately 260 bp; lane 5: DNA extracted from international standard samples of Aspergillus flavus URM7326; lane 6: Aspergillus terreus URM 7309; lane 7: Aspergillus fumigatus URM 7315; lane 8: Candida albicans (ATCC 52A); lane 9: Candida albicans (ATCC 18804); lane 10: Exophiala dermatitidis URM 7460; lane 11: Cryptococcus gattii URM7359; lane 12: Fonsecaea pedrosoi URM3161; lane 13: Paracoccidioides brasiliensis (Pb18); lane 14: Paracoccidioides brasiliensis Pb01.