TRUPCR® MTHFR Mutation Kit

The N5, N10-methylenetetrahydrofolate reductase (MTHFR) is an enzyme that plays an important role in the metabolisation of the amino acid methionine. Genetic polymorphism commonly associated with severe MTHFR deficiency is defined by a C to T substitution at position 677 (C677T) and A1298C defined by a A to C substitution at position 1298 of the MTHFR gene, which leads to the incorporation of amino acid alanine (A) instead of valine (V) at position 222 and glutamate to alanine substitution at codon 429 respectively of the MTHFR protein. The altered MTHFR is known as “Thermolabile MTHFR”. Homozygous and heterozygous carriers of these mutations both show reduced MTHFR activity. In particular, homozygous carriers suffer from significantly increased blood levels of homocysteine. In general, increased homocysteine levels are considered a risk factor of vascular diseases (e.g. arterial thrombosis) . The C677T mutation in its homozygous form alone or as a compound heterozygote, which involves both C677T and an A1298C condition (where an Adenine (A) residue changes to a Cytosine (C) residue at the 1298th position) lead to the disruption of the MTHFR gene and causes a drastic reduction of the MTHFR enzyme. This in turn, leads to an elevation of Homocysteine in the blood. The TRUPCR® MTHFR Kit is an allelic discrimination real-time PCR assay for qualitative detection of the mutation. In an allelic discrimination, two different probes specific for each allele are included in the PCR assay. Each probe is labelled with a different fluorescent dye (such as FAM or HEX/VIC) at its 5’ end and contains a non fluorescent quencher at the 3’ end. During qPCR amplification of the target DNA the probes will compete for binding across the variant region.