Background: Burden of pulmonary tuberculosis (PTB) is associated with success of directly observed therapy (DOT) services provided to patients suffering from PTB visiting a Tertiary Care Hospital in Aurangabad Dist., Maharashtra, India. Objective: This study was conducted to assess the outcome of repeat sputum AFB smear examination in patients who were smear negative on initial assessment in a Tertiary Care Hospital. Methods: Sputum AFB smears of suspected PTB patients done at RNTCP lab from January 2015 to December 2018, at MGM Medical college and Hospital were assessed. Results: Of the total 7043 PTB suspected patients examined in four years, sputum smear positive–PTB (SSP–PTB) cases were found to be 1095 (15.54%). A total of 321 cases were sent for repeat sputum examinationinsymptomatic initial sputum negative cases failing the antibiotic trial turned SSP–PTB were 62 (19.31%). Thus a considerable number of cases were missed in the initial screening (5.36%). Conclusion:A significant number of apparently smear negative TB cases may in fact be smear positive due to variousreasons and can be detected by a simple repeat sputum examination. Yield of sputum positive cases in repeat sputum examination is slightly higher as in initial sputum examination i.e. 19.31%. Therefore, the policy of repeat sputum examination in symptomatic initial sputum negative cases failing the antibiotic trial should be meticulously followed as advocated in the RNTCP diagnostic algorithm. Although the distribution of sputum positive smear results had no significant yearly variations and the overall burden of SSP cases did not decrease, therefore a lot of work needs to be undertaken on the PTB prevention and control program for the national action plan 2017– 2025.