This study was designed to investigate antifungal susceptibility and cross resistance of C. albicans isolates toward four antifungal drugs; Amphotericin B, nystatin, fluconazole and ketoconazole The study groups included 208 patients( 108 women and 100 children) who attented the Al-Suwayra Hospital/ Kut Province during the period from December 2008 to April 2009. Patients groups were represented by women who were complaining of vaginal discharge and itching and children who were complaining of oral thrush and infected with different type of disease include upper and lower respiratory tract infection, gastroenteritis, kala-azar and septicemia. The study showed that the infection percentage of vaginal candidiasis was 35.1% while oral thrush was 70%. The yeast isolates in two groups were C. albicans (76.8%), C. krusei (1.85%), C. tropicalis (5.5%), C. glabrata (11.1%), C. dublinances (0.9%), Trichosporon (1.85%) and Cryptococcus neoformans (1.85%). The activity of certain antifungal agents were tested against C. albicans isolates by using minimal inhibitory concentration (MICs). The results of range of MICs for amphotericin B was ( 8-128μg/ml), ketoconazole (0.5- 4μg/ml), fluconazole (8-64μg/ml), Nystatin (0.03-2μg/ml). The MICs of amphotericin B to the isolate that mutant to Nystatin tended to rise in parallel from (0.5-1μg/ml) to (4-16μg/ml). There was no correlation between MICs of these strains with fluconazole and ketoconazole. In addition, The MICs of ketoconazole to the isolates that mutant to fluconazole tended to rise in parallel from (0.5-2μg/ml) to (1-8μg/ml). There was no correlation between MICs of these strains with Nystatin and amphotericin B MICs. All the MICs values of the mutant strain return to the original values after remove the drug effect except the Nystatin . The MICs value for the isolate mutant to Nystatin dropped from the range (600-1200μg/ml) to (200-800μg/ml) but not returned to the original range (64-128μg/ml).