Molecular methods identification of dermatophytes using (PCR) techniques for patients in Al-Anbar governoret- Iraq

This study aims to isolate and diagnosis the Dermatophytes by mycological examinations methods in the laboratory, and identify the species that have appeared using the internal transcribed spacer (ITS) region and comparing the current local isolates with those provided by the National Center for Biotechnology Information (NCBI). Dermatophytes are predominantly caused by a group of closely related genera of Trichophyton, Microsporum, and Epidermophyton. These groups of fungi invade the stratum corneum of the skin, hair, and nails. Samples were collected from 100 patients with Dermatophytes in Ramadi Training Hospital, Anbar-Iraq. A portion of each sample was examined microscopically and the remaining portion of each sample was cultured onto plates of Saboraud Dextrose Agar added Chloramphenic and Gentamicin (SDACG). The results showed that out of the 100 cases of dermatophytes, only 60 (60 %) cases were positive by both direct KOH examination and culture, whereas false negative results was recorded in 20 (20 %) of specimens. Culture results of specimens showed nine types of dermatophytes were identified, n amely Trichophytonrubrum (26.0%), Microsporum canis (21.7%), T.mentagrophytes and their percentage (19.5), T. interdigitale (13.0%), T. ferrugineum (8.6%), T.soudanese (4.3%), and the rest of the three remaining species were Epidermophyton floccosum, T. tonsurans, T. verrcosum (2.2%). The most common clinical type of infection was tinea corporis (60.9%), and the others were as follows: Tinea cruris (19.6%), T. pedies (6.5%), T.faciei (6.5%), T. capitis (4.3%), and T. manus (2.18%). Females (52%) were more affected than men (40%). The countryside (65.2%) was more affected than the city (34.8%). The most affected age group was (31-40) years. Dermatophyte isolates were identified by studying the macroscopic and microscopic characteristics of their colonies. To ensure the correctness of their diagnosis, the primer (region ITS gene rRNA 18S) were used for the diagnosis of dermatophytes. The Polymerase Chain Reaction (PCR) products of Dermatophytes isolates were submitting to Macrogen Company for sequencing. Sequencing analysis showed nine new local strains recording in NCBI with accession numbers MW811374.1, MW811375.1, MW811376.1, MW811377.1, MW811378.1, MW811379.1, MW811380.1, MW811381.1, and MW811382.1. The genetic compatibility between the reference strain and the local strain was 99%. Keywords: Dermatophytes, Epidemiology, PCR