Spoligotyping and RFLP-IS6110 techniques were used to analyze 353 clinical Mycobacterium tuberculosis strains isolated from patients with tuberculosis in the Central Region of Russia. All spoligotypes were classified according to the international database. Most clinical M. tuberculosis strains in the collection obtained from patients living in the Central Region of Russia belong to the LAM (49.6%) and Beijing (34%) families, as shown by spoligotyping, and to A1 (51.3%) and W (34%) families, as evidenced by the RFLP-IS6110 technique.