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Introduction: Free –living Amoeba (FLA) are microorganisms that are abundant in the nature. Some genus of these protozoa are opportunistic and cause granulomatous encephalitis and lung infections in immunocompromised people such as cancer patients. Due to the little information about the status of infection of these parasites in the above patients, the present study was performed to determine the prevalence of FLA in cancer patients in Kashan hospitals. Materials and Methods: This cross-sectional study was performed on hundred oral cavity mucosal samples of cancer patients referred to Shahid Beheshti and Yathribi hospitals in Kashan during 2019 to 2020. The samples were cultured onto 1.5% Non-nutrient agar enriched with killed Escherichia coli and examined for the presence of FLA. The rate of contamination with free living amoebae and its relationship with demographic information of patients were analyzed by SPss. 16 Results: Overall, the prevalence of free-living amoeba in oral cavity of cancer patients was 89%. Also, 92.6% of breast cancer and 90.9% of bowel cancer patients were positive for FLA, respectively. The highest rate of FLA contamination was observed in women over the age of 66 years. In addition, the highest rate was observed in 90.9% of illiterate people and the lowest in 76.9% of academics, but the difference was not significant. Conclusion: The rate of free-living amoeba in oral cavity of cancer patients in Kashan were more than the study results of Iran and world. Therefore, health education about transmission and sanitation is recommended for prevention of infection.