Among the fungi, dermatophytes are the major cause of spectrum of superficial mycoses medically known as dermatophytosis (tinea) in human and animal. Treatment of these infections has still remained difficult. The aim of this survey was to evaluate in vitro anti-dermatophytic activity of ethanolic extract (EtOH) from Satureja khuzestanica leaf (SKLE) against some clinically important dermatophyte species from the genera of Trichophyton, Microsporum and Epidermophyton. Minimal inhibition concentration (MIC) of SKLE was tested against 14 dermatophyte strains of 5 species by using agar dilution method. Phytochemical screening of SKLE was carried out by High Performance Thin Layer Chromatography (HPTLC). The results of in vitro anti-dermatophytic activity of SKLE showed with MIC values between 1.250 and 10mg/mL. MIC90 and MIC50 values were as 0.625–1.250 and 0.156–0.312mg/mL, respectively. The MFC values of SKLE were in the range of 1.250–2.50mg/mL and possessed biological activity against dermatophytes. Morever, phytochemical analysis by HPTLC revealed that the ethyl acetate (EtOAc) extracts of SKL contain triterpenes which are known to have biological activity and it seems that this compound be responsible for the anti-dermatophytic activity of this plant. In conclusion, the results of in vitro antifungal susceptibility testing and phytochemical screening revealed that SKLE had both fungistatic and fungicidal activities against dermatophytes and can potentially be helpful as a supplementary or alternative for treatment of dermatophytosis.