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
Technology
  • Oligomix Factor II+V Duplex LT
  • instructions for use

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

Short informations
Sample
DNA from blood
Devices
LightCycler® 1.x
LightCycler® 2.0
LightCycler® 480
Duration
ca. 1,5 h
Determinations
20 reactions
100 reactions
Annex

The Instructions for use and the Material safety data sheet can be accessed via the login area. This area is conserved for Attomol customers. Please log in with your customer number and the corresponding password. For further informations please contact us directly.