Methylenetetrahydrofolate reductase (MTHFR) is an important enzyme involved in folate metabolism, which is essential for DNA synthesis and methylation. Genetic variations in the MTHFR gene seem to contribute to a decreased activity of MTHFR, ultimately confer increased susceptibility to stroke. To assess the association between this polymorphism and stroke risk, we conducted a comprehensive meta-analysis based on 73 eligible studies. A total of 73 studies, including 10,225 cases and 13,800 controls identified between 1999 and 2012, were selected through researching the PubMed, MEDLINE, EMBASE, Cochrane Library, Web of Science, and Chinese Biomedical Chinese National Knowledge Infrastructure and Literature database databases. Odds ratios (ORs) with corresponding 95 % confidence intervals (CIs) were used to assess the association. Overall, a significant elevated risk of stroke risk was associated with the rs1801133 polymorphism in all genetic models (homozygote model: OR 1.296, 95 % CI 1.109–1.514; dominant model: OR 1.179, 95 % CI 1.058–1.315; recessive model: OR 1.209, 95 % CI 1.063–1.375; allele comparison model: OR 1.154, 95 % CI 1.061–1.256). In the stratified analyses, significantly increased stroke risks were indicated among Asians in all genetic models (homozygote model: OR 1.726, 95 % CI 1.314–2.267; dominant model: OR 1.535, 95 % CI 1.282–1.838; recessive model: OR 1.452, 95 % CI 1.160–1.818; allele comparison model: OR 1.403, 95 % CI 1.211–1.626).The present meta-analysis suggests that rs1801133 polymorphism contributes to the risk of stroke, of note, in Asian populations.