China carries a heavy burden of cervical cancer (CC) and substantial disparities exist across regions within the country. In order to reduce regional disparities in CC, the government of China launched the National Cervical Cancer Screening Program in Rural Areas (NCCSPRA) in 2009. Critical to the success of the program are the health care workers who play a pivotal role in preventing and managing CC by encouraging and motivating women to use screening services and by providing identification and treatment services. This study aimed to assess cervical cancer knowledge among these health care workers at the county level in maternal and child health (MCH) hospitals across different socio-economic regions of China.A cross-sectional survey was conducted and self-administered questionnaires were sent to all health care workers (a total of 66) providing cervical cancer screening services in 6 county level MCH hospitals in Liaoning, Hubei and Shaanxi provinces, representing eastern, central and western regions of China; 64 (97.0%, 64/66) of the workers responded. ANOVA and Chi-square test were used to compare the knowledge rate and scores in subgroups.The knowledge level of the respondents was generally low. The overall combined knowledge rate was 46.9%. The knowledge rates for risk factors, prevention, clinical symptoms, screening and diagnostic tests and understanding of positive results were 31.3%, 37.5%, 18.1%, 56.3% and 84.4%, respectively. Statistically significant differences in scores or rates of CC knowledge were seen across the different regions. The total and sectional scores in the less developed regions were statistically significantly lower than in the other regions.The majority of the health care workers who provide CC screening service in NCCSPRA at county level MCH hospitals do not have adequately equipped with knowledge about CC. Given the importance of knowledge to the program's success in reducing CC burden in rural women in China, efforts are needed to improve the knowledge of health care workers, especially in less developed regions.