In the quest for natural drug products development, the antifungal activities of fractions of the leaf extract of Mitracarpus villosus S.W (D.C) (MV) against some fungal isolates were evaluated using reference drugs namely, ketoconazole, clotrimazole, fluconazole, nystatin and tioconazole. Leaves of MV were extracted successively with methanol, chloroform and ethyl acetate by cold maceration. The susceptibilities of solutions of the fractions in DMSO, (well diluted with sterile distilled water), and those of the reference antibiotics, certain mould and yeast isolates were tested using agar broth dilution method. Subsequently, the minimal inhibitory concentrations (MICs) were determined. Synergism of the fractions and the reference drugs was determined using the Checker-board method. All the fractions and their combinations with the reference drugs showed significant antifungal activities against the organisms used, with the chloroform fraction being the most active. The IZDs against Aspergillus niger were 15, 35, 27, and 9 mm for the methanolic extract, chloroform, methanolic and ethyl acetate fractions, respectively. The MICs obtained for the chloroform fraction were: 1.56, 6.25, 3.13, 6.25, 1.56, 3.13 and 1.56 mg/ml for the Aspergillus niger, Penicillium spp, Malassezia furfur, Aspergillus fumigatus, Candida albicans 1, 2 and 3 isolates, respectively. The highest synergism was observed for the chloroform fraction and ketoconazole combination (ratio 9:1) against Penicillium spp, at FIC index of 0.21. However, ketoconazole alone showed the highest activity against A. niger, Penicillum spp., A. fumigatus and all the candida isolates compared to the extracts/fractions, while clotrimazole showed the most activity against M. fufur. Based on the study, crude extract and fractions of MV leaves, especially the chloroform fraction, can be used alone or in combination with the tested antifungal drugs for treatment of superficial mycosis and candidiasis caused by susceptible organisms. Further characterization and development of standard dosage form of MV extracts is under focus.