Dermatophytosis is among the most common superficial mycoses in Iran. The purpose of this report was to update the clinical and mycological features of human dermatophytosis in the Khuzestan, southwestern Iran. In the framework of a one-year survey, a total of 4120 skin, hair and nail samples obtained from the outpatients with symptoms suggestive of tinea were analyzed by using direct microscopy, culture and molecular identification methods. Strains isolated from cultures were subjected to amplification of the nuclear rDNA ITS regions in a PCR assay followed by an early established RFLP analysis. For confirmation of species identification, 100 isolates as representatives of all presumable species were subjected to ITS sequencing. Infection was confirmed in 1123 individuals (27.25 %) in the age range of 1-89 years by direct microscopy and/or culture including 603 males versus 520 females. Frequencies of infections were the highest and the lowest in age groups of 21-30 and 11-20 years, respectively. Tinea corporis was the most prevalent clinical manifestation followed by tinea cruris, tinea capitis, tinea manuum, tinea pedis, tinea unguium, tinea faciei and tinea barbae. Trichophyton interdigitale (58.7 %) was the most dominant isolate followed by Epidermophyton floccosum (35.4 %), Microsporum canis (3 %), T. rubrum (1.5 %), T. species of Arthroderma benhamiae (0.5 %), T. tonsurans (0.3 %) and T. violaceum (0.3 %). Other species included M. gypseum, M. fulvum and T. verrucosum (each one 0.1 %). Such a high occurrence of infection with T. interdigitale, which has not been reported from Iran, is due to the use of accurate molecular methods based on new species concept in dermatophytes. The prevalence of dermatophytoses caused by zoophilic species remarkably increased and Trichophyton species of A. benhamiae has emerged as a new agent of dermatophytosis in southwestern Iran, while infections due to anthropophilic species, except E. floccosum, took a decreasing trend.