A biphasic culture system, the MB Check, was compared with conventional culture on Löwenstein-Jensen egg (LJ) solid medium and with Bactec broth culture for primary isolation of mycobacteria from clinical samples. A total of 104 mycobacterial isolates was detected from 985 samples examined by the three methods. The most sensitive primary isolation was with LJ culture and MB Check; these methods detected 93% and 87% of all positive cultures, respectively. MB Check allowed a somewhat more rapid detection than LJ culture. The presence of atypical mycobacteria was demonstrated most rapidly with the Bactec system.