Tuberculosis (TB) patients on the National Tuberculosis Elimination Program (NTEP) treatment protocol receive daily doses without health professional-supervised drug intake as during the previous directly observed treatment short-course (DOTS) regimen. We aimed to measure the level of adherence to anti-tubercular treatment (ATT) and the reasons for non-adherence among drug-sensitive TB patients on a daily-dose regimen in South India. A cross-sectional study was conducted among TB patients who received ATT as part of the standard treatment protocol in NTEP. Patients were interviewed to capture their understanding of TB, adherence, and the reason for non-adherence to ATT using validated instruments. Urine drug metabolite testing was performed using the high-performance liquid chromatography (HPLC) technique to confirm adherence. A total of 488 patients were recruited for the study. 64.8% of patients had 'good knowledge' about TB and ATT. According to the subjective report, 63.7% of patients were adherent, but urine drug metabolite testing revealed 53.4% adherence. A statistically significant difference (p< 0.05) exists between subjective and objective adherence measures. Patient-reported reasons for non-adherence were side effects of ATT (18.6%), loss of daily wages (15.0%), and forgetfulness (10.0%), among others. Nearly half of the patients were non-adherent to the daily dosing regimen. Adherence as reported by the patients is unreliable, and urine testing could be used in routine care to assess adherence. CTRI/2020/04/024941.