MTHFR Multiplex PCR Assay
The MTHFR mpx RealFast™ Assay is a fast and accurate multiplex real-time PCR test for the simultaneous detection of the 677C>T and 1298A>C mutations in the methylenetetrahydrofolate reductase (MTHFR) gene. The mutations are associated with decreased enzyme activity, which leads to hyperhomocysteinemia and to toxic side-effects of methotrexate therapy. The kit is designed to identify patients suspected to have an increased risk for cardiovascular diseases or intolerance to methotrexate. The qualitative assay discriminates the three possible genotypes for each allele in a human DNA extract. Reference sequence: NG_013351.1 g.14783C>T and g.16685A>C; dbSNP: rs1801133 and rs1801131.
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This kit in detail
The MTHFR mpx RealFast™ Assay is a fast and accurate multiplex real-time PCR test for the simultaneous detection of the 677C>T and 1298A>C mutations in the methylenetetrahydrofolate reductase (MTHFR) gene. The mutations are associated with decreased enzyme activity, which leads to hyperhomocysteinemia and to toxic side-effects of methotrexate therapy. The kit is designed to identify patients suspected to have an increased risk for cardiovascular diseases or intolerance to methotrexate. The qualitative assay discriminates the three possible genotypes for each allele in a human DNA extract.
Reference sequence: NG_013351.1 g.14783C>T and g.16685A>C; dbSNP: rs1801133 and rs1801131.
- Dual mutation detection: 677C>T and 1298A>C.
- Comprehensive coverage: full MTHFR gene analysis.
- Genotype differentiation: three alleles identified.
- Clinical impact: cardiovascular and methotrexate risk.
Elevated levels of homocysteine, an intermediary product of the methionine metabolism, are an established risk factor for atherosclerosis and arterial thrombosis. Hyperhomocysteinemia ultimately leads to endothelial dysfunction with associated platelet activation and thrombus formation. Mild to moderate forms can be caused by homozygosity for a common 677C>T point mutation in the coding region of the MTHFR gene. The second common MTHFR mutation 1298A>C also contributes to reduced enzyme activity, especially when occurring simultaneously with 677C>T. Fasting homocysteine levels are significantly higher in individuals heterozygous for both substitutions compared with individuals carrying only the heterozygous 677C>T mutation. Hyperhomocysteinemia may affect methotrexate sensitivity and contribute to toxicity, as MTHFR is an important enzyme in maintaining cellular folate pools. Thus, homozygosity for 677C>T or compound heterozygosity for 677C>T/1298A>C conveys a significantly higher risk for negative side-effects of methotrexate medication.
- RealFast™ 2x mpx Probe Mix
- MTHFR mpx Assay Mix
- MTHFR mpx WT-Control
- MTHFR mpx MUT-Control
The RealFast™ 2x mpx Probe Mix comprises HotStart Taq DNA polymerase and dNTPs in an optimized buffer system. The MTHFR mpx Assay Mix consists of MTHFR gene-specific primers and four allele-specific, dual-labeled hydrolysis probes. Controls representing wild type (WT-Control) and homozygous mutant (MUT-Control) genotypes are supplied with the kit.
The kit contains reagents for 100 / 32 reactions in a final volume of 20 µl each.
Datasheets and regulatory documentation
Everything your quality team needs for evaluation and procurement.
Instructions for Use
Frequently paired with this kit
Other assays and instruments lab teams add alongside the MTHFR Multiplex PCR Assay.
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