We investigated the HLA-DRB1*1501 tagging rs3135388 gene polymorphism and its association with multiple sclerosis (MS) susceptibility, disability and gender differences. The study group consisted of 306 MS patients and 137 healthy individuals. A significant difference in genotype distribution (Pg=3.06×10(-9)) and allele frequency (Pa=6.08×10(-10)) between MS patients and controls was demonstrated. The homozygotes AA and heterozygotes GA were more frequent in MS patients (OR=4.27, 95% CI: 2.64-6.92). A significant difference between female MS patients and female controls in genotype distribution (Pg=1.3×10(-8)) and allele frequency (Pa=2.82×10(-9)); (OR=5.11, 95% CI: 2.86-9.15) was also proved. Our results indicate that the distribution of the rs3135388 gene polymorphism is a risk factor for MS susceptibility in the Czech female population.