The Mycobacterium tuberculosis complex (MTBC) consists of species and subspecies that cause tuberculosis (TB) in humans and animals. As all MTBC strains are closely related at the molecular level, the exact distinction between them is not implemented in routine diagnostic techniques but could be used for tracking the zoonotic infection route, identification of drug resistance and epidemiological profiling. This study aimed to analyse the distribution of MTBC species in TB patients in the urban setting of Warsaw (central Poland). A total of 58 clinical isolates derived from patients diagnosed with TB were analysed. All specimens were processed for culture, DNA extraction and PCR. The distribution of species was investigated using GenoType MTBC molecular assay. All TB cases analysed were caused by Mycobacterium tuberculosis. M. tuberculosis appears to be the predominant cause of TB in Warsaw, central Poland, as no cases found of human TB caused by M. bovis or other MTBC strains were found.