Background Addressing HIV-related stigma among healthcare workers is vital in the era of contemporary antiretroviral therapy (cART). It is equally important for stimulating early diagnosis, and for meeting the complex medical needs of people living with HIV (PLHIV). Material and Methods. We analyzed the results of anonymous local survey comprised of 18 closed-choice questions on the knowledge and attitude towards HIV infection and PLHIV. The survey was conducted among 91 Bulgarian healthcare workers as a part of a large cross-national study in Europe and Central Asia, launched by ECDC and EACS in 2023. Results. The respondents were predominantly women (65,7%) working mostly as medical doctors (34%), 23% - in a specialized unit for PLHIV, 42% - with over 20 years of experience. The most important results were: lack of training on PrEP (in 62%), on stigma and discrimination (in 75%). Low level of knowledge on HIV transmission and prevention (in 40%) was associated with anxiety, exaggerated preventive measures and no professional experience with PLHIV. As compared to the mean EACS survey results, specific for Bulgaria were the high prevalence of the misconception “HIV is a result of irresponsible behavior, the ignorance of PrEP, the low rate of administrative sanctions related to PLHIV discrimination, and the low awareness of the availability of PEP. Conclusions There is an urgent need of targeted and tailored educational programs on HIV-related issues among the different healthcare workers groups. Those should be combined with legislative and administrative measures to assure the implementation of UNAIDS 2030 goals.