Abstract: In the present study, conventional bacterial isolation, bacteriophage-based assay (FAST Plaque TB) and Polymerase Chain Reaction (PCR) were evaluated for detection of Mycobacterium tuberculosis complex. A total of 47 mycobacterial isolates consisting of 38 isolates of Mycobacterium tuberculosis complex and 9 isolates of mycobacteria other than M. tuberculosis complex were used in this study. In addition, nine reference strains of Mycobacterium consisting of 7 Mycobacterium tuberculosis, one strain of Mycobacterium flavescens, one srtrain of mycobacterium duvalii were also used in this study. Conventional isolation is laborious, cumbersome and time consuming where it takes as long as 2 months for definitive diagnosis. The phage assay is sensitive and it takes only two working days. PCR is a rapid assay and definitive diagnosis of tuberculosis infection could be made possible within the same working day. The described bacteriophage-based assay could be used as rapid, sensitiveand specific method to support the currently available conventional methods used for detection of Mycobacterium tuberculosis in developing countries.