<b>Objectives</b><br/> Multidrug-resistant tuberculosis (MDR-TB) is a global problem that many countries are challenged with. Rapid and accurate detection of MDR-TB is critical for appropriate treatment and controlling of TB. The aim of the present study was to evaluate the TaqMan allelic discrimination without minor groove binder (MGB) as a rapid, efficient, and low-cost method for detection of drug resistant strains of <i>Mycobacterium tuberculosis</i>.<br/><b>Methods</b><br/> A total of 112 <i>M. tuberculosis</i> isolates from cases with diagnosed TB were subjected to drug susceptibility testing (DST), using the proportion method. Resistant isolates were tested for characterization of mutations in the <i>rpoB</i> and <i>KatG</i> genes by TaqMan genotyping.<br/><b>Results</b><br/> Of 112 <i>M. tuberculosis</i> isolates for which DST was performed, three, one, and two isolates were MDR, rifampin (RIF) resistant, and isoniazid (INH) resistant, respectively. According to the threshold cycle (Ct) and curve pattern of mutants, TaqMan probes detect all of the mutations in the analyzed genes (<i>katG</i> 315, AGC→ACC, <i>rpoB</i> 531, TCG→TTG, and <i>rpoB</i> 531, TCG→TGG).<br/><b>Conclusion</b><br/> The present study suggests that drug-resistant strains of <i>M. tuberculosis</i> can be detected by pattern’s curve or Ct with TaqMan probes without MGB in real-time polymerase chain reaction (PCR).