Methylenetetrahydrofolate reductase (MTHFR) is a gene involved in folate metabolism which may contribute to the risk of Alzheimer disease (AD). In this paper, an association of MTHFR-C677Tsingle nucleotide polymorphism (SNP) with the AD was evaluated by optimized PCR-confronting two-pair primers (PCR-CTPP) genotyping method. For this aim at first, PCR-CTPP method was optimized for detection of this SNP by additive agents and touchdown PCR. Then, an association of the mentioned SNP with the AD was assessed in a case-control study comprising 113 Alzheimer’s patients and 123 controls. Genotyping results showed that additive agents and touchdown PCR are two appropriate strategies for optimization of CTPP method. Our casecontrol study indicated a significant association between CT genotype (OR = 1.78, 95%CI = 1.04–3.03, p = 0.0357) and T allele (OR = 1.79, 95%CI = 1.07–2.10, p = 0.0273) with AD. Also, MTHFR-C677T polymorphism is associated with AD in a dominant model (OR = 1.79, 95%CI = 1.07–2.10, p = 0.0273). These results recommend that MTHFR-C677T polymorphism increases the risk of Alzheimer disease and can be a biomarker in the screening of this disease. And genotyping of aforementioned SNP could be performed by optimized PCR-CTPP a time-saving and reliable method.
