The global incidence of tuberculosis (TB) in prison is 5 to 70 times greater than that in the general population. The systematic screening and treatment of TB in prison benefit TB control and treatment outcomes. However, no study has supported this in Thailand. Here, we evaluated the effectiveness of long-term, continuous, systematic screening for TB control. We conducted a retrospective study in Suratthani Central Prison, Thailand, using annual TB screening from 2016 to 2020. A positive TB screening was defined as having any TB symptoms or chest radiography abnormality. We collected sputum smear and Xpert® Mycobacterium tuberculosis/rifampicin (MTB/RIF) results from those inmates with positive screening results. Bacteriologically confirmed TB was defined as a positive sputum smear or M. tuberculosis detected by Xpert assay. Inmates with clinically diagnosed TB were also eligible for this study. Those inmates diagnosed with active TB were sent to an isolation zone in prison and received anti-TB medicine using directly observed therapy and were monitored by pulmonologists during the treatment course via telemedicine. Of 18,965 TB screening events, 175 patients were diagnosed with TB. The 5-year prevalence was 922 per 100,000 inmates. The relative reduction in TB prevalence was 60.5% from 2016 to 2020. Overall, 94.3% of TB patients were successfully treated (sum of patients cured and treatment completed). Three TB patients died during the 5-year study period. All occurred in 2016. The systematic screening and treatment of TB in inmates effectively reduced the prevalence and mortality in Suratthani Central Prison. The success rate of treatment in this prison has been consistently high.