Genome-wide association studies provide information implicating 5,10-methenyltetrahydrofolate synthetase (MTHFS) as a candidate gene for renal disease. In the Atherosclerosis Risk in Communities study, the intronic single-nucleotide polymorphism rs6495446 in the gene MTHFS confirmed the association between this gene and renal disease among Caucasian participants. We replicated this genetic association in a Taiwanese population with diabetic nephropathy (DN). A total of 358 Taiwanese patients with type 2 diabetes (T2D) were recruited. The case group comprised 180 T2D patients with DN, and the control group comprised the remaining patients without DN. rs6495446 in MTHFS had no significant effect on the risk of DN in Taiwanese patients with T2D. Multivariate logistic regression analysis demonstrated that being male, the duration of diabetes, plasma triglyceride level, and glycemic control were factors that predicted the development of DN.