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Eswatini has the highest age-standardized incidence and second highest mortality rate related to cervical cancer globally. In Eswatini, community health workers educate communities about cervical cancer screening. They need to have accurate knowledge about cervical cancer and screening to do this effectively. Therefore, the current study aimed to assess knowledge regarding cervical cancer screening among community health workers in Eswatini. A telephone survey of 172 community health workers from eight selected constituencies was conducted. Descriptive analyses were performed to assess participants' socio-demographic and service-related characteristics. Linear regression was applied to investigate factors associated with cervical cancer screening-related knowledge. One hundred and seven (62%) participants answered at least 80% of the questions correctly. However, knowledge regarding cervical cancer risk factors, the meaning of screening results and Eswatini cervical cancer screening guidelines was suboptimal. Community health workers aged 46–55 were more likely (β = 1.27, 95% confidence interval [CI]: 0.39–2.15, p < 0.01) to have a higher cervical cancer screening knowledge score than those aged 30–45 years. Community health workers from Lubombo were marginally less likely (β = −0.83, 95% CI: −1.80 to −0.04, p = 0.05) to have a higher knowledge score than those from Hhohho. This study suggests knowledge deficits amongst community health workers in Eswatini. Knowledge deficits may result in inaccurate information being communicated to clients. While increasing knowledge of these vital health workers may not be sufficient to increase cervical cancer screening rates in Eswatini, it is an essential first step that should be the focus of future educational efforts. [ABSTRACT FROM AUTHOR]