Dermatophytoses are worldwide distributed superficial mycoses that affect the skin, nails and hair and are caused by keratinophilic fungi of the genera Trichophyton, Microsporum, and Epidermophyton. HIV-infected Patients are more susceptible to several fungal infections including dermatomycosis, which can present a gamut of clinical manifestations. Currently, several molecular methods have been applied in order to improve their diagnosis, identify their etiological agents and to improve the fungi taxonomy. Aim: To evaluate epidemiological and molecular aspects of dermatophytes isolated from HIV-infected patients at the Teaching Hospital in Uberaba, Minas Gerais State, Brazil. Methods: A total of 306 HIV-infected patients were included from august 2014 to december 2015. Samples of skin, hair and nail were collected from patients with or without cutaneous lesions. As control group, 92 HIV-negative patients with cutaneous lesions were evaluated. The isolated dermatophytes were phenotypically characterized and   molecularly identified by RFLP and ITS region sequencing. Results: Of the 306 HIV/AIDS patients, 153 (50%) presented cutaneous lesions that permitted yield several fungal species of which 22 (14.38%) were phenotypically identified as dermatophytes. From the control group were also obtained 19 isolates.  Trichophyton rubrum was the most common isolate species and the toenails were the body sites most affected in both groups. There was no statistical difference between the frequencies of dermatophytoses in HIV/AIDS patients with lesions compared to the control group. Although it is the gold standard of the mycological diagnosis of dermatophytoses, in the present study, the phenotypic identification of the isolates presented several difficulties. A total of 41 dermatophytes were submitted to molecular characterization. By RFLP technique were identified 28 (68.29%) of dermatophytes at the species level. The profiles obtained were variable among the different species, which allowed the visual differentiation between T. rubrum, Trichophyton interdigitale, Microsporum canis and Trichophyton verrucosum. The obtained patterns optimized the identification of three previously characterized isolates only at the gender level. The RFLP of the ITS region also allowed the correct identification of two T. rubrum isolates previously mistakenly named T. interdigitale. The amplified fragments generated sequences of approximately 630 bp, which allowed confirming the phenotypic identification in 70% of the ten sequenced isolates. The ITS region, although relatively conserved, was able to detect intraspecific variations among the dermatophytes and sequences obtained showed high identity with in relation to clinical isolates from different geographic regions deposited on GenBank. Multiple sequence alignment demonstrated the presence of two major clusters, corresponding to the complexes of T. mentagrophytes and T. rubrum species. Conclusion: Molecular techniques were complementary to the phenotypic diagnosis and contributed to the improvement of the correct species identification, however, phenotypic identification in most places is still considered the classic method of mycological diagnosis of dermatophytes at the clinical level.

Epidemiological and molecular aspects of dermatophytes isolated from HIV-infected patients in a Brazilian Teaching Hospital