ArticleMolecular biology reports2026
Development and validation of a tetra-primer ARMS-PCR assay for genotyping the MTHFR (rs1801131) c.1286 A > C (p.Glu429Ala) polymorphism: a comparative study with KASP.
Article in Molecular biology reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundThe tetra-primer amplification refractory mutation system (T-ARMS) PCR assay is a cost-effective and rapid method for SNP genotyping. The methylenetetrahydrofolate reductase (MTHFR) c.1286 A > C (rs1801131) polymorphism can influence MTHFR enzyme activity and homocysteine levels. While its independent clinical impact remains a subject of debate, it is frequently screened in thrombophilia panels, particularly in the context of compound heterozygosity. METHODS AND
resultsThis cross-sectional validation study aims to develop and validate a T-ARMS-PCR assay on the MTHFR c.1286 A > C mutation. The results were validated using the Kompetitive Allele-Specific PCR (KASP) as the reference method across 30 clinical DNA samples from a thrombophilia-susceptible cohort. The T-ARMS-PCR assay successfully distinguished genotypes with a clear electrophoretic separation and without non-specific amplification. The results exhibited 100% concordance with the KASP assay (95% CI: 88.4-100%; κ: 1.0). The total turnaround time including the reaction and electrophoretic separation was 107.5 min.
conclusionsT-ARMS-PCR offers a reliable, rapid, and cost-effective alternative for MTHFR c.1286 A > C genotyping. This method is particularly suitable for resource-limited clinical settings that require accurate genotyping without the need for high-end sequencing or specialized instrumentation.
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