Purpose: The role of healthcare workers in cervical cancer prevention is pivotal because healthcare workers’ recommendations have been reported to be a key motivational factor for cervical cancer screening among women in Nigeria. Assessing their knowledge, attitudes, and practices on cervical cancer prevention is necessary. Patients and methods: This is a cross-sectional survey involving healthcare workers in Ekiti State, Nigeria. A questionnaire designed for the study was used to obtain data about the knowledge, attitudes, and practices of the healthcare workers. Logistic regression was used to assess the determinants of practice and a p - value of less than 0.05 was taken as statistically significant. Results: Out of 188 participants that completed the study, 165(90.7%) had good knowledge about the prevention of cervical cancer while 24(13.2%) had good practice concerning the prevention of cervical cancer. positive attitude (Odds ratio 1.24, 95% CI 1.14 - 1.35, p < 0.001) increased the odds of good practice. Conclusion: There was no association between the knowledge of cervical cancer prevention and the practice of the healthcare workers. Further research is required to explore the incongruence between knowledge and practice of cervical cancer prevention and its practice.