The Microbiology and Clinical Microbiology Department, Gulhane Military Medical Academy, Ankara, Turkey, a tertiary referral hospital in a region endemic for tuberculosis. To evaluate rifampicin resistance of Mycobacterium tuberculosis complex strains using FASTPlaqueTB-RIF, a rapid and novel bacteriophage-based susceptibility technique. Results of isolates tested with the BACTEC 460 TB system were compared with FASTPlaqueTB-RIF. Susceptibility to rifampicin of M. tuberculosis complex isolates was tested for 88 isolates using FASTPlaqueTB-RIF. Sixty-seven isolates were susceptible and 21 were resistant to rifampicin using the BACTEC 460 TB system. Overall accuracy for FASTPlaqueTB-RIF was 94.3% (95%CI 87.3-97.5) for the detection of rifampicin susceptibility. The sensitivity and specificity of FASTPlaqueTB-RIF were respectively 100.0% (95%CI 84.5-100) and 92.5% (95%CI 83.6-96.7). This study demonstrates that FASTPlaqueTB-RlFM is a rapid and inexpensive test which has a good correlation with the BACTEC 460 TB system.