HIV and tuberculosis (TB) co-infection remain the leading causes of mortality in many developing countries. Several policies for their prevention and management exist; however, progress to meet the World Health Organization's indicators is slow. As nurses in rural areas are the main health care professionals responsible for the implementation of policies, questions arose about nursing knowledge regarding TB and HIV policies. A concurrent mixed method research approach was used to investigate this issue in a rural area of the Western Cape in South Africa. After a two-step sampling process, data were collected through 44 questionnaires and 12 interviews. We used descriptive and inferential statistics as well as content analysis methods. We found that most of the nurses had insufficient knowledge on certain aspects of policy guidelines that included medications, intensive case findings, and management of multidrug-resistant TB. It is recommended that training be intensified to improve implementation of policies.