Background: There are active and latent tuberculosis types, a global public health concern. Test confirmation is required to distinguish between active tuberculosis and nontuberculous mycobacterial infection. This study aims to evaluate the relationship between the type of lower airway secretion specimen, Mycobacterium tuberculosis positivity rate of MGIT-LJ, as well as the relationship between the Mycobacterium tuberculosis positivity rate in MGIT-LJ and radiological findings. Methods: This study is cross-sectional with a prospective design. The sample is the result of the examination of the MGIT-LJ method and medical record data of patients suspected of Pulmonary TB at Dr. Soetomo Academic Hospital Surabaya Indonesia that meets the inclusion criteria, analysis of the culture results of MGIT-LJ method and radiological findings were carried out to assess the relationship between the character of sputum, the positivity rate of Mycobacterium tuberculosis and radiological findings. Result: A total of 137 from June 2022 till November 2022 found 88 (64.2%) with spontaneous sputum, 6 (4,4%) aspirate ETT, and 43 (31.4%) BAL. The positivity rate of the MGIT-LJ culture method of the spontaneous sputum specimen to detect Mycobacterium tuberculosis was 20.5% higher than ETT or BAL specimens because the implementation of clinical indication to request MGIT-LJ culture method of ETT and BAL specimens is still confusing. The specificity of the chest radiography diagnostic test from all specimens type of 61.86%, with a weak accuracy rate. Conclusion: In this study, the positivity rate of the MGIT-LJ culture method of the spontaneous sputum specimen to detect Mycobacterium tuberculosis was 20.5% higher than ETT or BAL specimens. Chest radiography has a weak accuracy level and must be complemented test with the culture method.