Introduction: The human immunodeficiency virus (HIV) affects mainly CD4+ T lymphocytes, debiliting the immune system. Guidelines and recommendations have changed due to the emergence of new prevention and treatment strategies. Pre-and post-exposure prophylaxis (PrEP and PEP) constitute prevention modalities in seronegative people based on the administration of antiretroviral drugs (ARV) with the aim of reducing the probability of transmission with high rates of effectiveness. General objective: Determine the level of knowledge about HIV, its prophylaxis and prevention in healthcare workers and health science students in Paraguay. Methodology: Observational, descriptive cross-sectional study, non-probabilistic sampling of consecutive cases, the questionnaire "Knowledge about HIV and pre-and post-exposure prophylaxis to HIV" was administered. Results: 504 people were surveyed with an average age of 29 +/- 7 years, 58.3% (294) being female, with 22% (111) from Central department. The level of knowledge about HIV prophylaxis and prevention was sufficient in 54.2% (273) of the participants, without statistically significant differences with respect to the specialty practiced. Conclusion: The level of knowledge was sufficient in just over half of the healthcare workers surveyed.