A total of 105 fresh sputum samples were collected from patients with negative acid-fast for pulmonary tuberculosis, which was obtained from the Thoracic and Respiratory diseases Treatment Center for Tuberculosis in Baghdad city. The samples were decontaminated, concentrated and cultured on Lowenstein (LJ) medium, incubated at 28-30°C and observed the growth after 3 days-4 weeks. The results indicated Non-tuberculous mycobacteria (NTM) were isolated 4/105 (3.8%) from the patient, and all of these isolates belong to RGM (rapidly growing mycobacterium) based on their ability to grow on LJ medium, cultural characteristics, acid-fast stain, and biochemical tests. At molecular identification, M. fortuitum 50% (2/4) was isolated from a male who was smoking for a long time, M. porcinum 25% (1/4) was isolated from diabetic females and the last isolate was M. cosmeticum 25% (1/4). All the patients were elderly patients aged 41-75 years; this is the first study of isolating these species from pulmonary infections in Baghdad city.