PCR Development for the Diagnosis of Dermatophytes in Equines

Abstract

Development and standardize of PCR for diagnose of most common equine dermatophytes was carried out. We used one reference strain and four isolates characterized by routinely methods from clinical samples. Sabouraud broth was used to obtain the fungal mass by long 8 days incubation. Once the mass was dried, it was pulverized with liquid nitrogen in a mortar and deposited in tubes to obtained DNA by phenolic extraction. NCBI GenBank data were used for the primer design, the sequences were aligned manually and once the initiators were selected, they were placed in the DNAMAN program, in order to differentiate the five species involved. We diagnosed successfully common dermatophytes in Equidae by standardized PCR test. A total of 50 samples were used for the challenge test, 22 have been declared positive by conventional diagnostic methods, and the remaining samples were selected randomly the obtained results were similar compared with conventional test. M. canis primers were highly sensitive. For the others species we need more samples to corroborate the usefulness of the test.