In the present study, 34 new pulmonary tuberculosis cases prediagnosed upon clinical and radiological findings, have been evaluated by means of microbiological aspects such as microscopy [Ehrlich-Ziehl Neelsen (EZN) and Auramin-Rhodamine (A-R) stains], culture [inoculation into Lowenstein-Jensen (LJ) media] and three different molecular methods [Cobas Amplicor Mycobacterium tuberculosis polymerase chain reaction (PCR), Gen-Probe amplified M. tuberculosis direct test and TaqMan Real Time ABI 5700 PCR]. M.tuberculosis positivity of the sputum samples were 55.8% (n: 19) with EZN staining, 52.9% (n: 18) with A-R staining and 58.8% (n: 20) with culture methods. Among molecular techniques, Cobas Amplicor and TaqMan PCR yielded positive results in 21 patients (61.7%), however, the positivity rate of Gen-Probe AMTD was 47.0% (n: 16). Of 34 samples, 10 (29.4%) were found to be positive with all of the methods used, while six (17.6%) of them were negative with all methods. It can be concluded that molecular methods may aid to the diagnosis of tuberculosis, in conjunction with clinical findings and bacteriological results.