Detection of C677T & A1298C mutations within the MTHFR gene by PCR and RFLP assays and assessment of risk factor of Hyperhomocysteinemia

The MTHFR gene within the human genome, codes for the synthesis of Methylenetetrahydrofolate Reductase enzyme, which reduces 5,10-Methylenetetrahydrofolate to 5-Methyltetrahydrofolate, which in turn, is the major circulatory form of folate in the blood. Folate, in this form, among it’s other functions, is involved in reducing the homocysteine levels in the blood, whose elevated levels lead to Hyperhomocysteinemia, causing various major disorders. Mutations within the gene lead to impairment of gene function, in turn causing the homocysteine levels to rise. The C677T and A1298C mutations are the main causative agents for MTHFR gene disruption. During the course of the project, a total of 79 samples were analyzed for the presence of these mutations. The blood samples were first subjected to PCR, giving two separate DNA fragments each responsible for either of the conditions. The fragments were then subjected to RFLP analysis to detect the mutations. The results were finally given with respect to the risk factor faced by each individual based on a molecular diagnostic point of view