Background Cervical cancer is a major global public health concern, with 85% of cases occurring in low- and middle-income countries. In South Africa, it is the second most common cancer amongst women. Screening and treatment of cervical cancer precursor lesions is associated with a lower incidence and mortality. This research determines the associations between women's beliefs about cervical cancer and screening and the uptake of Papanicolaou (Pap) smears in Johannesburg, where cervical screening uptake is suboptimal. Methods This research was approved by the University of Witwatersrand Human Research Ethics Committee (Medical), clearance certificate number: M170243 and the Johannesburg District Heath Research Committee prior to conducting the study. All participants signed a consent form prior to participating in this study. This cross-sectional analytical study used an interviewer-administered validated measurement scale based on the Health Belief Model (HBM) to describe health beliefs regarding cervical cancer and screening among 280 women aged 30 years and older, attending Johannesburg primary care facilities in 2017. Logistic regression models, with robust estimation of variance to account for clustering of women within clinics, were fitted to identify health beliefs (perceived susceptibility, severity, barriers and benefit, cues to action, and self-efficacy) associated with ever having had a Pap smear (screening uptake), while controlling for knowledge of screening and potential confounders. Results Of the 280 women, 177 (63.2%) had ever been screened, 180 (64.3%) were never married, 199 (71.1%) attained secondary education and 133 (47.5%) were employed full time. Women of older age (AOR = 1.6 for a 5-year increase in age; CI: 1.3-1.9; P Conclusions This study shows that women who take up screening are older, more knowledgeable regarding cervical cancer and screening, less likely to perceive screening barriers, and more likely to perceive cervical cancer as a severe disease. This highlights that for public health interventions to increase screening uptake, the focus should include tailored behaviour change communication strategies that address women's beliefs regarding screening barriers and emphasize the severity of cervical cancer.