TB diagnosis and resistance detection remain to suffer from complex techniques and long time taking. The rapid identification of drug-resistant strains of Mycobacterium tuberculosis is crucial for the timely initiation of appropriate antituberculosis therapy. Over the years, several new technologies have been proposed to accelerate and simplify the detection of resistant M. tuberculosis. In this context, we evaluated the Genotype MTBDRplus to detect resistance to isoniazid and rifampin in positive M. tuberculosis cultures. The performance of the Genotype MTBDRplus assay was compared with that of the phenotypic drug susceptibility tests, a gold standard culture based method. The Genotype MTBDRplus assay was quicker and more cost-effective for the detection of rifampin and isoniazid resistance, with a slightly lower detection of rifampin-resistant strains in our setting.