Background: Drug resistant tuberculosis and Multi drug resistance TB are the major global public health threat causing high morbidity and mortality in many parts of the world. Different risk factors are associated with the emergence of drug resistant tuberculosis. A public health response towards these risk factors is important to cut the line of TB transmission. Data on the full drug susceptibility testing profiles of M. tuberculosis isolates to first line drugs and associated factors among pulmonary tuberculosis and tuberculosis lymphadenitis are limited in the study area. Objective: The main aim of this study was to assess the first-line anti-TB drug resistance profile and associated factors among pulmonary tuberculosis and tuberculous lymphadenitis cases in selected health facilities of Bahir Dar city, Northwest Ethiopia. Materials and Methods: A facility-based comparative cross-sectional study was conducted among 160 PTB and TBLN patients from May 01 to August 30, 2022. Data on sociodemographic and clinical factors were collected using a pre-tested questionnaire. The sputum and fine needle aspirate and 5 ml of blood samples were transferred to sterile falcon and cryo tubes and serum separator tube respectively, then transported to Amhara Public Health Institute using triple package. The samples were inoculated in to liquid (Mycobacterium growth indicator tube, (MGIT Broth) and solid (Lowenstein Jensen, (LJ) media. First-line anti-TB drug resistance profile was determined from freshly grown isolates by convectional phenotypic DST methods using BACTEC-MGIT 960 machine. Data were entered and analyzed using IBM SPSS statistics for windows version 26. Descriptive statistics and logistic regression analysis were computed. Finally, data were summarized using texts, tables, percentages, figures and inferential statistics. Results: The overall prevalence of DR-TB rate was 15/160 (9.37%). Among those, most of DR-TB cases were PTB 12 (7.5%) and the rest 3 (1.87%) were TBLN cases. When looking individual drugs, the rate of isoniazid, rifampicin, ethambutol, pyrazinamide and streptomycin resistance were 11(6.87%), 7(4.37%), 6 (3.75%), 4 (2.5%) and 1(0.62%), respectively. Relatively, high rate of mono drug resistance was observed to INH, 4 (2.5%) followed by PZA, 3 (1.8%). The proportion of INH, RIF, EMB and PZA poly drug resistance was 1(0.62%) and when excluding PZA, the poly drug resistance (INH+RIF+EMB) became 4 (2.5%). The prevalence of MDR-TB was 7(4.37%), among these 5 (3.12%) were from PTB. Conclusions and recommendations: TB lymphadenitis not considered as a special reservoir for DR-TB when compared with PTB. The prevalence of drug resistance among new TB patients is fairly stable over the years. However, the high resistance rate of INH over RIF pose challenge to management of DR-TB and latent TB treatment. Occupancy, alcohol drinking, contact history, housing ventilation, cough persistent for more than two weeks, high level of triglycerides and forms of TB showed significant association with DR-TB in the present study. Health education on the role of isolation of TB cases, Housing ventilation by opening windows, and avoidance of living on congregate setting must be given to cut the TB transmission chain. Further study on the prevalence of drug resistance among TBLN cases and role of TBLN in DR-TB should be addressed. Keywords: tuberculosis, first line anti TB resistance, MDR-TB, PTB, TBLN, Risk factors.