Aim: Tuberculosis is a chronic, necrotizing disease known since prehistoric times. The most important stage of the tuberculosis control program is the microbiological identification, typing and detection of drug resistance of the Mycobacterium tuberculosis complex for the detection of active cases. The aim of this study is to compare the results of the methods used in the detection of M.tuberculosis complex in clinical samples. Materials and Methods: The results of the samples sent to the laboratory for the purpose of investigating M. tuberculosis complex between January 2016 and January 2022 from patients with pre-diagnosis of tuberculosis were evaluated retrospectively. In microbiological diagnosis, Ehrlich-Ziehl-Neelsen (EZN) staining method, culture (BACTEC MGIT 320 automated system and Löwenstein Jensen (LJ) medium, and polymerase chain reaction (PCR) tests were used. Results: A total of 1706 samples with all three requests (EZN, culture and TB-DNA) were included in the study. The mean age of the patients was 46.49±25.77 years and 1025 (60.1%) were male. Culture in 48 (2.8%) samples, PCR positivity in 40 (2.3%) and EZN positivity in 32 (1.9%) samples were mostly sent from the departments of chest diseases (52.8%) and pediatrics (24.1%). Conclusion: It was determined that the culture method gave more reliable results than PCR and EZN in demonstrating the presence of M. tuberculosis complex. As a result, since the culture method, gives late results, extensive routine use of molecular tests such as PCR is needed for accurate diagnosis of tuberculosis in a short time and to detect drug resistance.