Antifungal resistance represents a major clinical challenge to clinicians responsible for treating invasive fungal infections due to the limited arsenal of systemically available antifungal agents. Currently, fungal invasive infections are related to antifungal resistance by pathogenic fungi. The current study assumed to use of natural ingredients such as the fruiting bodies of large fungi, specifically Agaricus gennadii. This study is also a new initial step in the field of producing an antifungal that has a promising and safe future. However, most of the used antifungals such as polyenes, azoles, echinocandins, and flucytosine have side effects and may lead to damage to the human body. To achieve the goal of this study, 120 mid-stream urine samples were collected from urinary tract infection (UTI) patients, who attended Samarra General Hospital, the primary care sector and some medical clinics in the city of Samarra/Salah al-Din during the period from 11-1-2020 to 1-2-2021. Many laboratory examinations were performed including: microbiological, biochemical, and molecular tests of 70 samples of UTI patients, which develop yeast colonies on culture plates and were considered as positive results. Our results showed that the extract of the Agaricus gennadii fruit bodies contained a number of organic compounds including phenolics, flavonoids, saponins, terpenoids and alkaloids. Regarding the susceptibility of the isolated yeast species, many concentrations (100%, 75%, 50% and 25%) of the fungal extract were investigated. Data analysis of the obtained results showed that among all tested yeasts, Trichosporon mucoides and Candida parapsilosis were susceptible to the fungal extract at all concentrations, however, no effect of the fungal extract on the rest of the studied yeasts. Also, our results demonstrated that the susceptibility was increased with the increase of the fungal extract concentration. More studies are needed to separate and test the exact role of these compounds in the inhibition of fungal growth.