Tuberculosis (TB) is a major worldwide health problem, particularly in India, where it accounts for a quarter of infections. Adherence to the directly observed treatment, short-course (DOTS), which is necessary for TB treatment, is difficult in resource-limited circumstances. This study addressed the knowledge, attitude, and perceived barriers related to TB and DOTS adherence among patients and caregivers. A cross-sectional survey was undertaken at the selected TB clinics between November 2023 and March 2024. Purposive sample yielded 180 patients and 217 caregivers. Data were obtained using validated, self-structured questionnaires that assessed knowledge, attitude, and perceived barriers to TB and DOTS adherence. Statistical analysis was carried out with IBM SPSS version 23.0. The study found that caregivers had considerably greater TB knowledge than patients (p<0.05). Many patients misunderstood the cause of TB (50.6%) and embraced various misconceptions about the disease, such as TB spreading by shaking hands or sharing food, swimming in a holy river to cure diseases, and TB being the result of sin or karmic retribution. Distance to treatment facilities (95.6%), the necessity to take time off from work (91.7%), and social stigma (65.0%) were identified as the top three barriers to DOTS adherence. The study highlighted considerable gaps in knowledge, distant healthcare facilities, busy work schedules, and social stigma as barriers to TB treatment adherence in Deoghar, Jharkhand. Addressing these challenges via focused education and support activities is essential for enhancing DOTS adherence and TB treatment outcomes.