A combination of two methods -- a rapid culture method [Mycobacteria Growth Indicator Tube (MGIT); Becton-Dickinson, USA] and a double polymerase chain reaction (PCR) assay -- was assessed for the detection and identification of Mycobacterium tuberculosis and Mycobacterium avium from clinical samples. The aim of the study was to evaluate the ability of the system to offer rapid and accurate diagnosis of mycobacterial infections. After decontamination, clinical samples (n = 554) were stained and cultured in parallel on solid media and in MGITs following standard procedures. The performance of the two culture systems was compared. Positive MGITs were tested for the presence of Mycobacterium tuberculosis and Mycobacterium avium by PCR of IS6110 (Mycobacterium tuberculosis) and the 16S rRNA gene (Mycobacterium avium). A total of 41 mycobacteria -- 27 Mycobacterium tuberculosis isolates, eight Mycobacterium avium isolates, and six other species of mycobacteria -- were isolated by one or both culture media. The MGIT system recovered 36 (87.8%) mycobacteria and the solid media 33 (80.4%). The mean time to detection by the two culture systems did not differ overall, but the mean time to detection of Mycobacterium avium from smear-positive specimens was shorter in MGITs than in solid media (5.25 days vs. 16.25 days, P < 0.05). The double PCR assay performed on the 36 positive MGITs correctly identified all 24 Mycobacterium tuberculosis-positive MGITs and all six Mycobacterium avium-positive vials. Therefore, application of the PCR assay to positive MGITs may mean that Mycobacterium tuberculosis and Mycobacterium avium can be identified at an earlier stage than with current methods.