The objective: to evaluate changes in detection frequency of Mycobacterium tuberculosis of the main phylogenetic lines with various genetic determinants of resistance to rifampicin, isoniazid and fluoroquinolones in 1998–2003 and 2016–2021.Subjects and Methods. 965 mycobacterial cultures were studied, those cultures were isolated from pulmonary tuberculosis patients in 1998–2003 and 2016–2021. The spoligotypes of isolated Mycobacterium tuberculosis cultures and presence of gene mutations associated with resistance to isoniazid, rifampicin, and fluoroquinolones were determined.Results. In 2016–2021 versus 1998–2003, the incidence of Beijing subline increased from 50,72% to 64,60%. Within Euro-American line, the detection frequency of subline LAM9 decreased (from 40,00% to 14,81%), while the detection frequency of sublineT1 increased (from 27,11% to 41,36%). Mycobacterium tuberculosis with MDR genotype rpoB531_Ser->Leu + katG315_Ser-> Thr [1] was selected. Mycobacterium tuberculosis with genotypic resistance to fluoroquinolones was detected only in cultures of 2016–2021, and Mycobacterium tuberculosis with preXDR genotype was significantly more common in Beijing subline compared to Euro-American line (21,67% vs. 3,73%, p <0,05).