No research has been conducted to explore the variables associated with healthcare providers’ (HCPs) knowledge and attitudes toward the human papillomavirus vaccine (HPV) since the vaccine was approved for free use in some Chinese cities. In Shenzhen, southern China, a convenience sample strategy was used to distribute questionnaires to HCPs involved in the government’s HPV vaccination program from Shenzhen. There were 828 questionnaires collected in total, with 770 used in the analysis. The mean HPV and HPV vaccine knowledge score was 12.0 among HCPs involved in the government HPV vaccination program (with a total score of 15). the average scores for HPV and HPV vaccine knowledge varied among different types of medical institutions. District hospitals had the highest mean score of 12.4, while private hospitals ranked fourth with a mean score of 10.9. Multivariate logistic regression results revealed significant disparities in the type of license and after-tax annual income across HCPs (p< 0.05). The future education and training for HCPs should focus on private community health centers (CHCs), HCPs whose license type is other than a doctor, and HCPs with low after-tax annual income.