Malawi is facing a severe HIV and AIDS epidemic with an estimated 12% of its population living with the virus. Health workers are on the front lines of the HIV epidemic and they face the risk of HIV infection in both their personal and professional lives. This mixed method study aimed to explore the enablers and barriers to HIV counselling and testing and antiretroviral therapy by health workers in Malawi. After qualitative data were collected through in-depth interviews with health workers in the Mchinji and Nsanje districts, a survey questionnaire was constructed and administered to 906 health workers in eight districts in Malawi. A majority (76%) of health workers surveyed reported having undergone HIV testing and counselling, of whom 74% reported repeat testing. A striking result of the study is that 22% of health workers reported testing after occupational exposure to HIV. The proportions of respondents reporting that they tested after experiencing symptoms, or self-testing for HIV were 11% each. The in-depth interviews and the survey revealed multiple challenges that health workers face to accessing HIV testing, counselling and treatment, including fear of a positive result, fear of stigma and lack of confidentiality. Additional barriers included health workers' personal acquaintance with those conducting testing, along with their perception of being "role models" which could exacerbate their fears about confidentiality. Given health workers' critical role in HIV delivery in Malawi, there is need to develop solutions to help health workers overcome these barriers.