Background: Several studies have established strong links between stigma and discrimination against PLWHA by health workers and poor utilization of HIV/AIDS prevention and care services by them. Aim: This study aimed to assess the attitudes and practices related to stigma and discrimination against persons living with HIV/ AIDS (PLWHA) among health workers in a tertiary care facility in Sokoto, Nigeria. Materials and Methods: This was a cross-sectional study among 258 health workers selected by systematic sampling technique. A structured self-administered questionnaire was used to collect data on the research variables. Data were analyzed using IBM SPSS version 20 statistical computer software package. Results: The mean age of the respondents was 32.9 ± 9.3 years; majority of them were males (52.3%), married (67.1%), and were nurses (67.1%). Majority of respondents showed negative attitudes (indicating stigma) towards PLWHA, as 74.0% would prefer to refer them to other physicians for treatment, and 67.4% were not comfortable with giving injections or performing invasive procedures on PLWHA. Large proportions of respondents had been involved in discriminatory practices against PLWHA such as testing patients for HIV without consent (41.1%), and administering a differential treatment for patients with HIV/AIDS (48.1%). Conclusion: Attitudes and practices related to stigma and discrimination against PLWHA were very prevalent among the respondents in this study. Management of hospitals, government and other stakeholders involved in HIV/AIDS prevention and care should ensure full implementation of the strategic interventions for eliminating stigma and discrimination against PLWHA to prevent a resurgence of HIV/AIDS epidemic across the country.