The bacteria Mycobacterium tuberculosis causes pulmonary tuberculosis (PTB), which is the biggest cause of infection-related mortality globally. Some socio-demographic characteristics, TB/HIV co-infection, co-morbidity contact history, and past treatment history status are the key risk factors for developing tuberculosis. The main objective of this study was to investigate the prevalence of Mycobacterium tuberculosis infection, Rifampicin resistance tuberculosis, and risk factors among presumptive pulmonary tuberculosis patients at Injibara General Hospital in Injibara town, Northwestern Ethiopia. A total of 422 randomly selected study participants were participated in this study. Accordingly, 422 sputum samples was collected and subjected for TB laboratory examination. In addition, a structured and pre-tested questionnaire was distributed to the selected study participants to determine the associated risk factors for Mycobacterium tuberculosis. Hospital based cross-sectional study design was used. All the collected data were analyzed by using SPSS version 25. Logistic regression analysis was done to determine the strength of association between dependent variables and risk factors. Variables with a p value of less than 0.25 during bivariate logistic regression analysis were selected for multivariate analysis. A p-value of less than 0.05 was used to indicate statistical significance. A total of 422 presumptive pulmonary tuberculosis patients were included. Mycobacterium tuberculosis was detected in 48 participants (11.7%). Of these participants, three cases (6.25%) were resistant to rifampcin. The prevalence of Mycobacterium tuberculosis infection was statistic ally significant association with age (AOR=4.43695%CI=1.495-13.168), poor ventilation status ( AOR=4.910; 95%CI=2.229-10.814), contact history with known tuberculosis patients (AOR=3.2 71; 95%CI=1.303-8.210), HIV co infection, and alcohol consumption (AOR=3.340; 95%CI=1.572-7.094). Prevalence of Mycobacterium tuberculosis and rifampicin-resistant M. tuberculosis was high. Age, living in poor ventilation status home, HIV co infection, contact with known PTB patients, and alcohol consumption were the major risk factors for pulmonary tuberculosis. Keyword; Ethiopia, Injibara, Mycobacterium tuberculosis, pulmonary tuberculosis, rifampcin resistance tuberculosis