Evaluation of a tissue factor dependent factor V assay to detect factor V Leiden: demonstration of high sensitivity and specificity for a generally applicable assay for activated protein C resistance.
Resistance to the anticoagulant effects of activated protein C (APC) is now considered the most prevalent cause of inherited thrombophilia. The great majority of patients with activated protein C resistance (APCR) have a missense mutation in the factor V molecule (factor V Leiden, FVR506Q) resulting in defective inactivation of factor Va due to a loss of an APC cleavage site. The diagnosis of APCR has been based upon the inability of APC to prolong the activated partial thromboplastin (aPTT) clotting time in subjects with APCR. However, this assay has a number of deficiencies which limit its general use. We have evaluated a newly described one-stage tissue factor dependent factor V coagulation assay for APCR in 117 patients and controls and compared the results of this assay in a blinded manner to a polymerase chain reaction (PCR) based assay for the molecular defect of factor V Leiden. 43% (50/117) of the patients studied were receiving coumadin or heparin, or had a lupus anticoagulant. The tissue factor dependent factor V assay had 100% specificity and sensitivity for factor V Leiden and successfully predicted a homozygous state in the three documented homozygotes. The PCR-based assay for factor V Leiden resulted in a single false positive assay due to a silent A to C transition at nucleotide 1692 resulting in the loss of the Mnl restriction endonuclease cleavage site. The single-stage tissue factor dependent factor V assay is a highly sensitive and generally applicable assay for APCR.
Liebman
H A
HA
Department of Medicine, University of Southern California School of Medicine, Los Angeles 90033, USA.
Sutherland
D
D
Bacon
R
R
McGehee
W
W
D001780:Blood Coagulation Tests / Q000379:methods* / Q000592:standards; D001796:Blood Protein Disorders / Q000175:diagnosis*; D005165:Factor V / Q000032:analysis*; D006801:Humans; D011486:Protein C / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013925:Thromboplastin / Q000032:analysis*
D011486:Protein C; C095381:factor V Leiden; D005165:Factor V; D013925:Thromboplastin