FV and PTH are major FV-PTHplayers in the coagulation cascade. The FV Leiden mutation causes a single amino acid exchange at position 506 (R506Q), which alters a cleavage site and thereby prevents efficient inactivation of FV. Persisting FV activity increases the risk of clot formation in veins. The PTH 20210G>A mutation in the 3’ untranslated region results in increased mRNA synthesis and higher prothrombin plasma levels, which in turn lead to elevated thrombin generation and consequently to excessive formation of fibrin clots.

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The FV-PTH mpx RealFast™ Assay is a fast and accurate multiplex real-time PCR test for the simultaneous detection of the two most important thrombophilic mutations. The point mutation 1691G>A in the human coagulation Factor V (F5) gene, referred to as FV Leiden, and the prothrombin (PTH) 20210G>A mutation in the Factor II (F2) gene are associated with hereditary thrombophilia. The kit is intended to test patients suspected of having an increased risk for thrombotic disorders. The qualitative assay discriminates the three possible genotypes for each of the mutations in human genomic DNA: normal, heterozygous or homozygous mutant.

FV-PTH mpx RealFast ™ RT-PCR Assays technology

The ViennaLab RealFast™ Assays are based on real-time PCR and hydrolysis probes, also commonly called TaqMan® probes. The sequence-specific oligonucleotide probes carry a fluorescent reporter dye at the 5´-end and a quencher dye at the 3´-end. While the probe is intact, the quencher is close enough to the reporter to suppress the fluorescent signal of the 5´-fluorophore. During the combined annealing/extension phase of PCR, the probe is cleaved by the 5´ to 3´ exonuclease activity of Taq DNA polymerase, thereby separating the fluorophore from the quencher dye. This process results in detectable fluorescence, which is proportional to the amount of accumulated PCR product.