Evaluation of Primers OPF-01, P54, and 1253 to Identify A. fumigatus, A. flavus, and A. niger from Polymorphic Patterns Obtained by RAPD-PCR

We evaluated the specificity of the primers OPF-01, P54, and 1253 to identify <i>A. fumigatus</i>, <i>A. flavus</i>, and <i>A. niger</i>, respectively, with the RAPD-PCR method. Eighty-two isolates belonging to the sections <i>Fumigati</i>, <i>Flavi</i>, and <i>Nigri</i> were used. The isolates were identified by phenotypic (macro- and micromorphology) and genotypic (partial sequences of the <i>BenA</i> gene) methods. The RAPD-PCR method was used to obtain polymorphic patterns with the primers OPF-01, P54, and 1253. The specificity of the polymorphic patterns of the isolates of each species was evaluated through the UPGMA clustering method and logistic regression model. All isolates of the genus <i>Aspergillus</i> were identified at the section level by macro- and micromorphology showing the typical morphology of the sections <i>Fumigati</i>, <i>Flavi</i>, and <i>Nigri</i>, and the species were identified by the construction of the phylogeny of the partial sequence of the <i>BenA</i> gene. The patterns' polymorphic strains obtained with the primers OPF-01, P54, and 1253 for the isolates of <i>A. fumigatus</i>, <i>A. flavus</i>, and <i>A niger</i>, respectively, showed the same polymorphic pattern as the reference strains for each species. To verify the specificity of the primers, they were tested with other species from the sections <i>Fumigati</i>, <i>Flavi</i> and <i>Nigri.</i> The results support that the primers OPF-01, P54, and 1253 generate polymorphic patterns by RAPD-PCR species specific to <i>A. fumigatus</i>, <i>A. flavus</i>, and <i>A. niger</i>, respectively.