Over recent years, cervical cancer incidence and related mortality have steadily increased in Eswatini. Low cervical cancer screening uptake partly explains the situation. Cervical cancer screening-related knowledge is positively associated with screening uptake. Little is known about women's cervical cancer screening-related knowledge in Eswatini. This study aimed to assess cervical cancer screening knowledge and associated factors among Eswatini women eligible for screening. A cross-sectional study involving three hundred and seventy-seven women aged 25 to 59 selected from four primary healthcare clinics in Eswatini was conducted. A paper and pen survey assessed knowledge about cervical cancer risk factors, benefits of screening, the meaning of screening results, recommended screening intervals, and socio-demographics. Descriptive analyses were performed to assess participants' sociodemographic characteristics. Linear regression was applied to examine associations between cervical cancer screening-related knowledge and participants' sociodemographic characteristics. Two hundred and twenty-nine (61%) participants answered 80% or more knowledge questions correctly. Compared to HIV-positive participants, HIV-negative participants had 0.61 times lower cervical cancer screening knowledge scores (β = -0.39, 95% CI: -0.56, -0.19, p = 0.03). Participants who travelled more than 30 minutes to the clinic had 0.3 times lower cervical cancer screening knowledge scores (β = -0.70, 95% CI: -1.15, -0.25, p < 0.01) compared to participants who travelled less than 30 minutes to the clinic. Relatively high overall cervical cancer screening knowledge levels were observed among the study participants. Findings from the current study may inform future educational programs to create and sustain an accurate understanding of cervical cancer screening in Eswatini communities.