attomol® Factor II+V Duplex Realtime LT

REF 1181

20 reactions

REF 1228

100 reactions

Intended purpose

The assay attomol® Factor II+V Duplex Realtime LT is used for the supportive diagnosis of thrombophilia. This test can be used for the simultaneous determination of the transitions 20210G>A in the human factor II gene (prothrombin, rs1799963) and 1691G>A in the human factor V gene (Leiden mutation, rs6025). It is a manual, qualitative real-time PCR assay based on a LoopTag probe, which is evaluated by a melting curve analysis. The sample material has to be genomic DNA prepared from EDTA or citrate blood.

Field of application

In the second-last step of the blood coagulation cascade, prothrombin (factor II) is converted into the serine protease thrombin. This converts fibrinogen molecules by splitting the firbrinopeptides A und B into fibrin monomeres, which spontaneously aggregate to fibrin polymers. The replacement of guanine (G) at position 20210 of the prothrombin gene by adenine (A) (factor II [prothrombin] mutation) causes an increase in the prothrombin concentration in the blood plasma and thus promotes the development of thromboembolism [Poort et al., 1996, Blood 88: 3698-3703]. The incidence of this mutation is 1-3 % in the European population. In carriers of the heterozygous mutation in prothrombin gene, the risk of thrombosis increases 3 to 5-fold compared with healthy people in combination with other factors (intake of oral oestrogen-containing contraceptives, surgical operations, pregnancy, increased age, smoking) [Markis et al., 1997, Thromb. Haemost. 78:1426-1429]. The factor V Leiden mutation, which has a prevalence of approximately 5 % in the European population, is one of the most important hereditary risk factors for thrombosis. In the blood coagulation cascade factor V forms a complex (prothrombin activator) together with the activated factor X, phospholipids and calcium ions. This complex converts prothrombin (factor II) into thrombin, which causes a fibrin polymerization. The inactivation of factor V occurs through the activated protein C (APC). The substitution of guanine (G) for adenine (A) at position 1691 of the factor V gene results in an amino exchange in the binding site for APC. This amino exchange leads to the fact that factor V can only be insufficiently cleaved and thus inactivated [Bertina et al., 1994, Nature 369: 64-67]. The resulting accumulation of factor V in the blood leads to an increased coagulation tendency, which results in a higher risk of thrombosis in the affected patients. Homozygous mutations in the factor V gene lead to a significantly higher risk than heterozygous defects [Zöller et al., 1999, Haematologica 84: 59-70].

Kit content
  • Oligomix Factor II+V Duplex LT
  • instructions for use

You can find more informations about the Looptag-Technology here.

Short informations
DNA from blood
LightCycler® 1.x
LightCycler® 2.0
LightCycler® 480
ca. 1,5 h
20 reactions
100 reactions

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