This study aimed to compare and analyze the results of microbiological and immunological laboratory investigations of pulmonary tuberculosis and non-tuberculosis patients from Hospitals at Saudi Arabia, and also to evaluate the serodiagnosis detection of tuberculosis promptly using immunological assays in comparison with the traditional Ziehl Neelsen microbiological techniques. This is to develop reliable, effective assay, shorter duration and more specific diagnosis of active tuberculosis. The sensitivity, specificity and efficiency of Ziehl-Neelsen stain were 48 %, 100 % and 64 %, respectively. An identified antigen in serum samples of infected persons with pulmonary tuberculosis was detected using TB-Spot ELISA technique. The sensitivity, specificity and efficiency of TB-Spot ELISA for the detection of antigen were 97 %, 95 % and 98 %, respectively. The serum levels of TNF-α reach statistical difference (P< 0.0001) in patients positive for TB antigen in comparison with the levels in negative individuals for TB.