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Updated: May 18, 2026
A Method to Study the C924T Polymorphism of the Thromboxane A2 Receptor Gene
Published on: April 1, 2019
Development and validation of a tetra-primer ARMS-PCR assay for genotyping the MTHFR (rs1801131) c.1286 A > C
Amany Alqosaibi1,2, Akram Husain Rehman Syed Rasheed3, Huseyin Tombuloglu4
1Department of Biology, College of Science, Imam Abdulrahman Bin Faisal University, Dammam, 31441, Saudi Arabia.
Background:
The 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 Results:
This 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.
Conclusions:
T-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.
