The gold standard assay for the factor V Leiden mutation
involves the use of polymerase chain reaction (PCR) methodology.
These samples were submitted to the laboratory for factor V Leiden mutation
detection, and the genotype was initially established by PCR followed by restriction fragment analysis based on the method described by Koeleman et al.
The factor V Leiden mutation
prevents proteolytic cleavage of factor V by protein C, which leads to resistance of activated protein C.
Additional genetic risk factors for venous thromboembolism in carriers of the factor V Leiden mutation
The Quest samples used in the evaluation of the photo-cross-linking assay had previously been tested for factor V Leiden mutation
status by a PCR-based assay that used the Read-It[TM] detection system (Promega) (7).
m]s of homozygous mutant DNA for the prothrombin 20210A and factor V Leiden mutations
Automated detection of factor V Leiden mutation
using the LCx microparticle enzyme immunoassay.
When testing is called for, he recommended testing for factor V Leiden mutation
and the prothrombin mutation and performing immunoassays for anticardiolipin antibodies (for antiphospholipid syndrome) and lupus anticoagulant.
Activated protein C resistance and factor V Leiden mutation
are independent risk factors for venous thromboembolism.
Studies conducted to date have revealed a number of modulator genes, including angiotensinogen, tumor necrosis factor-alpha, glutathione S-transferase P1, lipoprotein lipase, HLA-G, methylenetetrahydrofolate reductase, factor V Leiden mutation
, plasminogen activator inhibitor-1, and insulin-like growth factor 11.
The assay uses oligonucleotide probes modified with photo-activatable crosslinker molecules (7) and has been used to detect the factor V Leiden mutation
The conventional procedures for mutation analyses were as follows: the factor V Leiden mutation
was detected by PCR followed by MnII restriction analysis and 3% Wide Range agarose electrophoresis as described (4); the prothrombin mutation was detected by the method of Poort et al.