Introduction: Cervical cancer screening helps detect precancerous lesions which when not treated can develop into invasive cervical cancer. Low prevalence of cervical cancer screening among female employees of Mulago Hospital prompted the researcher to assess the factors influencing uptake of cervical cancer screening among female employees of Mulago Hospital. Methodology: A cross sectional study that targeted female employees of Mulago Hospital was done using a self-administered questionnaire. Data analysis was done using SPSS software version 20, presented in frequencies and percentages; and Chi Square test was used to test association of factors and uptake of cervical cancer screening. Results: 64.1% of respondents had ever screened for cervical cancer. The socio demographic characteristics that showed significant association to cervical cancer screening uptake were age and nature of employment. Cervical cancer awareness was high among all respondents. High levels of adequate knowledge, positive attitude and good cervical cancer practice was reported more among Nurses/Midwives, Doctors and paramedical staff than among administrative and support staff; and were significantly association to cervical cancer screening uptake. Good practice of cervical cancer was found among 60% of respondents and there was strong significant association to all hospital factors that was assessed. Conclusion: Uptake of cervical cancer screening was higher among female employees of Mulago Hospital compared to the national average and it was higher among Nurses/Midwives, Doctors, and paramedical staff than administrative and support staff. Recommendation: Ministry of Health and Mulago Hospital should develop and implement national cervical cancer program that targets all eligible women, including female staff of Mulago Hospital. In addition, they should harmonize cervical cancer screening data nationally to inform planning and implementation of cervical cancer prevention program.

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