In South Africa, the prevalence and death rates as a result of cervical cancer remains high, creating social and economic instability. The main objective of this study was to determine factors affecting participation in cervical screening by female nurses in public health institutions in Vhembe district, Limpopo Province. Early diagnosis and treatment are essential in cervical cancer screening as the prevalence of the disease decreases. The study was carried out at public health institutions in Vhembe district, Limpopo Province. A quantitative, descriptive, cross-sectional design was used in this study. Structured self-reported questionnaires were used in the collection of data. Descriptive statistics were used when analysing data to identify statistically significant differences in variables using SPSS version 26, and the findings were presented in percentages to generate evidence for the study. According to the study findings, (218, 83%) female nurses had been screened for cervical cancer, while the minority (46, 17%) had not been screened. The reasons cited were that they thought they were healthy (82, 31%), (79, 30%) felt embarrassed, and (15%) feared positive results. The majority (190) of them had last been screened more than three years before, with only a few (27, 10%) screened within the previous three years. A hundred and forty-two (53.8%) displayed negative attitudes and practices towards screening if it is paid for, and a hundred and eighteen (44.6%) perceived themselves as not vulnerable to acquiring cervical carcinoma. Furthermore,(128, 48.5%) strongly disagreed and 17(6.4%) were undecided about being screened by a male practitioner. The study concluded that negative attitudes, poor perception, and embarrassment are factors leading to low uptake by female nurses. Therefore, this study recommends that the Department of Health should build the capacity of nursing staff on matters of national significance to achieve sustainable goals and be a healthy nation. Nurses should be at the forefront of departmental programmes.