Background: Poor Physician Adherence to DOTS strategy is one of the reasons for the failure of tuberculosis control. The aim of this study was to evaluate the level of adherence of physicians in Birjand to DOTS and determine the related factors. Methods:This research was an analytical cross-sectional study. The study population consisted all general physicians, internal physicians, pediatrics and infection disease specialists who were active in Birjand in 2024. A valid and reliable questionnaire, including demographic and 20 scenario-based multiple-choice questions based on the DOTS strategy was used to collect data. Differences in compliance among groups were assessed using T-test and ANOVA, and factors associated with compliance were determined using multiple linear regression analysis with SPSS version 26. Results:All eligible physicians in study were 121 in which 109 individuals responded to the questionnaire (response rate: 91%). The mean age was 37.28 ± 7.5 years, 56% of them were male and 75.2% (82 persons) were general practitioners. Approximately 15% of physicians had no familiarity with DOTS. The mean adherence score to DOTS was 10.51 ± 4.5 out of 20 which rated as inappropriate. The adherence scores in diagnosis, follow-up, and follow-up of the contacts were 2.42 ± 1.07, 1.99 ± 1.22, and 2.61 ± 1.06 out of 4, while for treatment was 3.49 (± 1.76) out of 8. Multiple linear regression analysis indicated a significantly higher adherence score among female compared to male physicians (β = 1.65, p = 0.02). Prior good familiarity (β = 5.4, p < 0.001) and brief familiarity with DOTS (β = 2.47, p = 0.019) were significantly associated with a higher adherence score compared to no familiarity. Conclusion:The adherence of Birjand physicians to the DOTS was inappropriate. Females and physicians with Prior good and brief familiarity with DOTS had a higher compliance rate.