An estimated global total of 10.6 million people had Mycobacterium tuberculosis infection, causing tuberculosis (TB) in 2022, equivalent to 133 cases per 100,000 people. Kenya remains a high-burden country for drug-susceptible TB, and human immunodeficiency virus (HIV), western Kenya bearing the highest burden of HIV. A 2016-2017 survey in Kenya demonstrated that the estimated burden of TB was 169,000 people against 85,188 cases diagnosed in 2016, implying more than 40% of adults were missing, and Siaya was among top 10 high TB burden counties in Kenya. For healthcare workers (HCWs), TB remains an occupational hazard, especially in countries with high TB burden, with HCWs having a 2-3-fold increased risk of developing TB compared with the general population. This cross-sectional study investigated the prevalence and occupational risk factors for TB among HCWs in Siaya County, using a sample size of 275 from the 10 level 4 hospitals. Sputum samples were collected from all participants for TB testing using Gene Xpert. Self-administered structured questionnaires and observation checklists were used to collect data from the HCWs. Out of the 275 respondents, 18 participants reported to have been diagnosed with TB in the previous five years, giving a period prevalence of 6.5%, while none of the participants who produced sputum for TB testing turned positive. Only 68 (24.7%) CHWs had been trained on TB, and health facilities that conducted health talks to patients at waiting bays and segregated coughers to be seen first were significantly associated with reduced TB occurrence among HCWs (ꭓ2 = 11.125, p = 0.012). Potential occupational risk factors such as constant supply of N95 masks (ꭓ2 = 7.505, p = 0.069), putting on N95 masks (ꭓ2 = 6.335, p = 0.135), provision of designated cough corner/sputum sample collection place (ꭓ2 = 6.714, p = 0.116), availability of functional infection prevention and control (IPC) measures (ꭓ2 = 7.796, p = 0.067), availability of TB health talk schedules (ꭓ2 = 8.353, p = 0.052), and availability of adequate lighting and cross ventilation in all waiting bays and clinical rooms (ꭓ2 = 6.065, p = 0.144) were not significantly associated with TB incidence among healthcare workers. Unavailability of certain IPC interventions was associated with the occupational risk of TB among healthcare workers. The Ministry of Health should enhance the dissemination of relevant TB manuals to update staff on IPC and availing N95 masks to all healthcare workers, while facilitating frequent TB screening to identify HCWs with TB, and all put on early treatment.