Abstract: ObjectiveThis study aims to explore knowledge, attitudes, behaviors of prevention of mother-to-child transmission(MTCT) of and discrimination against human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) among medical staff in order to promote elimination of MTCT.MethodsRelevant medical staffs were selected from midwifery institutions using convenient sampling method. “Questionnaire Star” a web-based electronic questionnaire were used to complete this survey. The content of the survey included socio-characteristics, knowledge about MTCT, and discrimination against HIV/AIDS. Differences of knowledge about MTCT, score of discrimination against HIV/AIDS scale were compared between various kinds of health professionals.Results1 273 health professionals were recruited. Their average age was 38. 6 ± 8. 8(21-64). 210 people answered all the questions correctly, accounting for 16. 5%. Among them, the percentage of doctor was higher than that of nurses(χ²= 11. 377,P= 0. 003);the percentage of health professionals with senior titles was higher than that of other staff(χ²= 9. 631,P= 0. 008); the percentage of medical staff who were first married was higher than that of people who were unmarried/divorced(χ²= 6. 949,P= 0. 008). The average scores of discrimination against HIV/AIDS Scale was 2. 10 ± 0. 55, 2. 35 ± 0. 86 for dimension 1(fear of infection) ranked first, followed by 2. 22 ± 0. 67 for dimension 2 (discrimination, blame and prejudice), 2. 17 ± 0. 69 for dimension 3 (social discrimination) and 2. 02 ± 0. 60 for dimension 4(personal discrimination). Multivariate analysis showed age, marriage, awareness and field of specialization were associated with scores of discrimination against HIV/AIDS Scale.ConclusionAwareness of preventing from MTCT of HIV/AIDS should be improved among medical staff. Worry about being infected is the main reason for discrimination.Increased awareness may help to reduce discrimination against HIV/AIDS.