Background: Healthy sexual function can be considered as an important element to improve personal and public hygiene. The sexual desire plays an important role in mental health and improving the quality of life. Objectives: The current study aimed to compare sexual function of females in urban and rural areas. Methods: The current descriptive study adopted 800 females of reproductive age (range 15 -45 years) referred to rural and urban healthcare centers in Ahvaz, Iran, in 2015. Samples were randomly selected. Applied instruments in the study were demographic information and female sexual dysfunction questionnaires (FSFI). Independent T-test, Chi-square and logistic regression were employed to analyze data by SPSS ver. 22. Results:The result showed a significant statistical difference between females in urban and rural areas in terms of sexual desire, vaginal lubrication, intercourse pain and sexual function (P < 0.05). However, there was no significant statistical difference between the two groups in terms of excitement, orgasm and sexual satisfaction (P > 0.05). Frequency of sexual dysfunction was 59.9% in females in rural and36.5% in urban areas and the difference between the groups was statistically significant (0.000). In both groups, the highest sexual disorder frequency was related to intercourse pain. Conclusions: According to the obtained results, females in the rural areas had lower sexual function than the ones in the urban areas. It is suggested to establish female sexual health units in healthcare centers to give female sexual function consultation adjusted with awareness and culture of females and consider the existing problems.