Objective To understand the current status of primary healthcare workers’ knowledge of hepatitis B and their willingness to engage in non-discriminatory behaviors， to explore the factors influencing their willingness， and to provide a basis for health education on hepatitis B for primary healthcare workers in Qinghai Province.

Methods A questionnaire survey was conducted among primary healthcare institutions in eight cities （prefectures） of Qinghai Province using a combination of random sampling and convenience sampling. The questionnaire included demographic information， knowledge related to hepatitis B， and willingness to engage in non-discriminatory behaviors.

Results A total of 393 subjects were included in the survey. The accurate awareness rate of hepatitis B transmission route among primary healthcare workers in Qinghai Province was 36.89%. Of the respondents， 46.31% were unwilling to live with family members infected with HBV or carriers， and 50.13% were unwilling to dine with friends infected with HBV or carriers. The willingness of primary healthcare workers to engage in non-discriminatory behaviors related to hepatitis B varied by different places of residence， gender， marital status and per capita household income （P<0.05）. Regarding different dimensions of HBV-related knowledge， there were significant differences in age， education level， and per capita household income for transmission route （P<0.05）. Vaccination showed significant differences based on marital status and per capita family income （P<0.05）. There were significant differences in treatment and outcome among different gender， education level， and per capita family income. Anti-discrimination policies varied significantly by place of residence， age， ethnicity， and per capita income. Multivariate logistic regression analysis showed that primary healthcare workers’ knowledge of hepatitis B transmission route was a significant factor influencing their willingness to engage in non-discriminatory behaviors （OR=0.49， 95%CI：0.29‒0.84）.

Conclusion It is essential to continuously strengthen hepatitis B health education for primary healthcare workers in western China to reduce the willingness to engage in non-discriminatory behaviors towards hepatitis B and protect the rights of HBV-infected individuals.