Objective: This study evaluated the efficacy of educational program on knowledge and practice of pap smear test among female healthcare workers in Al Falah & Banyas healthcare centers, UAE Design: Pre-test post-test quasi-experimental study was conducted on efficacy of educational program on knowledge and practice of pap smear test Methods: 108 female health workers were selected using convenience sampling in Al Falah & Banyas healthcare centers, UAE. Data was collected using a self-administered structured questionnaire, analyzed by SPSS version 25. Results: 42.6% work in Al Falah, while 57.4% are in Baniyas healthcare center. The pre-education program shows that 80.6% of female health workers had low and middle knowledge scores regarding cervical cancer and 78.7% about a pap smear test. Post-training, all the study participants had good knowledge scores regarding cervical cancer (p=0.001) and pap smear tests (p=0.030). Poor knowledge score was found among young female health workers (p=0.030) and fewer years of experience (p=0.011) in health workers, physicians, and nurses (p=0.000). Conclusion: The study showed gaps in knowledge among female health workers regarding cervical cancer and Pap smear test. In-service training program about cervical cancer and its screening is an effective educational method to provide an effective base for cervical cancer prevention and improve female health workers’ knowledge and uptake. Practice guarantees quality of life of women