Background: Tuberculosis is one of most dangerous and infectious diseases that widespread in the world, as untreated severe cases lead to the death of infected patients. Objective: The current study aimed to isolate and diagnose Mycobacterium tuberculosis and to know the number of infections in the city of Samarra (Iraq), and whether there are associations between sex or age and disease. Patients and methods: A total of 50 specimens were collected from TB patients, out of which 25 were males and 25 were females. The diagnosis was made by the physician at the center based on chest x-rays, microscopy, biochemical tests and examinations. The confirmatory examination was carried out using a PCR test.  Results: The age of the participants ranged from 20 to 45 years. The diagnosing bacteria from the number of the samples reached 123 samples collected from people suffering from chest symptoms who arrived at the Respiratory Disease Center in Samawah-Muthanna Governorate. The results of isolation and diagnosis by bacterial cultures, x-ray showed and PCR that 49 (39.9%) samples were for people with tuberculosis and 72 (60.1%) samples were not infected with the disease. The liver enzymes were found to be significantly low compared to the normal levels (P<0.05) in the studied patients. The number of the persons who completed the treatment and were cured of the sickness was discovered to be 24 (about 75%); we considered the importance of the moral difference (P value 0.463), because the significance level is larger than 0.05. Conclusions: High proportion of drug resistance to M. tuberculosis strain detected that could suggest the need to increase the efforts to the strengthen TB control program in the study area. There were significant variations observed in the drug resistance patterns between M. tuberculosis lineage.