The study aimed to identify Trichoderma harzianum isolates morphologically and by using PCR teqnique and evaluate the antifungal activity of T. harzianum CA-07  crude extract against Trichophyton mentagrophytes and Microsporium canis from patients with dermatophytosis. T.harzianum isolates were collected from the soil of Baghdad University gardens and they were identified depending of morphological features on plate and microscopic examination. The genomic DNA of T.harzianum isolate was extracted at a final concentration of (400-600) μg / 2-3 g of wet mycelium and at a purity of 1.6-1.8and DNA sample was amplified with each of universal primers (ITS-1& ITS-4) to be used for detection of Trichoderma species. The crude extract was extracted from T.harzianum strain CA-07 by ethyl acetate with finalyield of 4.8gm., then different crude extract concentrations (0.5, 1, 2 and 4 mg/ml) were used against the clinical pathogenic fungi (T.mentagrophyte and M.canis) using agar well diffusion method. The results of identification of Trichoderma harzianum isolates by using PCR confirmed that the isolate was T.harzianum strain CA-07,and their crude extract exhibited significantly high antifungal activity against T. mentagrophyte and M. canis with high growth inhibition zones(14, 12mm) respectively at the lower concentration (0.5mg/ml)of crude extract.