Background: Tuberculosis (TB) is recognized as an important health risk for health workers, however, the absence of occupational health surveillance has created knowledge gaps regarding occupational infection rates and contributing factors. This study aimed to determine the rates and contributing factors of active TB cases in laboratory healthcare employees at the National Health Laboratory Service (NHLS) in South Africa, as identified from an occupational surveillance system. Methods: TB cases were reported on the Occupational Health and Safety Information System (OHASIS), which recorded data on occupation type and activities and factors leading to confirmed TB. Data collected from 2012 to 2019 were used to calculate and compare TB risks within NHLS occupational groups. Results: During the study period, there were 92 cases of TB identified in the OHASIS database. General workers, rather than skilled and unskilled laboratory workers and medical staff, had the highest incidence rate (422 per 100,000 person-years). OHASIS data revealed subgroups that seemed to be well protected, while pointing to exposure situations that beckoned policy development, as well as identified subgroups of workers for whom better training is warranted. Conclusions: Functional occupational health surveillance systems can identify subgroups most at risk as well as areas of programme success and areas where increased support is needed, helping to target and monitor policy and procedure modification and training needs.