Background Tuberculosis is one of the infections targeted for eradication and is still under surveillance. Hospital workers in Nigeria take little or no precautions against tuberculosis while attending to patients. Objectives This research aimed to study the prevalence of the reactivity of positive Tuberculin Skin Tests among healthcare workers in a Nigerian University Teaching Hospital. Methods Ethical approval and informed consent were obtained from the participants. Two hundred and ten consented participants were recruited for this study, while 185 contributed till the end. The Tuberculin Skin Test was conducted following the standard operating procedures. The chi-square test was used to compare the variable at a significant level of P < 0.05. Of those who received a BGC vaccine < 3 years, 4.8% were excluded from the analyses. Results Mean ± SD of participants’ age was approximately 30 ± 8 years. Participants comprised 87 (47%) males and 98 (53%) females. The modal age group was 21-25 years. Most participants were clinical medical students, 93(50.3%), while doctors, 7 (3.8%), were second to least. The prevalence of positive reactivity to the Tuberculin Skin Test was 29.9%. There was no significant difference observed regarding sexes and positive reactivity. A significant difference was found in reactivity ≥ 10 mm between those who didn’t receive BCG and those who received BCG vaccine ≥ 10 years before the study. Conclusion Healthcare workers can be potential active TB patients and reservoirs. We recommend highly sensitive and specific periodic TB screening, prophylactic treatment, and re-vaccination of Health Care Workers in Nigeria.