BACKGROUND፡ Mycobacterial infections are known to cause a public health problem globally. The burden of pulmonary disease from nontuberculous mycobacteria is reportedly on the rise in different parts of the world despite the fact that there is limited data about the disease in sub-Saharan Africa including Ethiopia. Hence, we aimed to assess the magnitude of M. tuberculosis and nontuberculous mycobacteria (NTM) among presumptive pulmonary tuberculosis patients attending St. Paul’s hospital Medical College, Addis Ababa, Ethiopia.METHODS: A cross-sectional study was conducted from June to September 20/2016. Morning sputum specimens were collected, processed and cultured in Lowenstein Jensen medium and BACTEC MGIT 960 media. The nontuberculous mycobacteria were further confirmed and characterized by Genotype CM/AS assays. The socio-demographic, clinical and chest x-ray data were collected using a structured questionnaire. The data was analyzed using SPSS version 20.RESULTS: Out of 275 presumptive tuberculosis patients enrolled in the study, 29(10.5%) were culture positive for Mycobacteria. Of these, 3(10.3%) were found to be NTM and 26(89.6%) were Mycobacterium tuberculosis complex. Of the NTM, two were unidentified and one typed as M.peregrinum. There was no coisolation of Mycobacterium tuberculosis complex and nontuberculous mycobacteria. Overall, 6(23.1%) Mycobacterium tuberculosis complex isolates were resistant to at least one antituberculosis drug. Of these, two were multidrug resistanttuberculosis cases (7.7%) detected from previously treated patients.CONCLUSION: Relatively low magnitude of Mycobacterium tuberculosis complex and nontuberculous mycobacteria isolates were seen in the study area. Therefore, further study using a large sample size is needed to be done to consider nontuberculous mycobacteria infection as a differential diagnosis in presumptive pulmonary tuberculosis patients.