Modification of the ProC® Global assay using dilution of patient plasma in factor V-depleted plasma as a screening assay for factor V Leiden mutation

The activated protein C (APC) resistant-factor V (factor V Leiden) has emerged as the most common inherited risk factor for thrombosis in the Caucasian population. Beside DNA analysis, the laboratory diagnosis is often based on the detection of a poor anticoagulant response to exogenous APC. The ProC((R)) Global assay (Dade Behring, Marburg Germany) is a global clotting assay, which was primarily developed to evaluate the functionality of the protein C anticoagulant pathway. It is based on the ability of endogenous APC, generated by activation of protein C by an extract from Agkistrodon contortrix contortrix venom, to prolong an activated partial thromboplastin time. It was previously found to be highly sensitive for the factor V Leiden mutation and for protein C deficiency, but only moderately sensitivity for protein S deficiency. Here, we evaluated the performance of a modification of the ProC((R)) Global assay using a 1:5 pre-dilution of patient plasma in factor V-depleted plasma in the screening of the factor V Leiden mutation-related APC resistance. For that purpose, we investigated selected frozen plasma samples from 341 patients with a history of venous thromboembolism. The sensitivity for the factor V Leiden mutation of the modified assay was found to be 100%, as all the carriers of that mutation (five homozygotes and 77 heterozygotes) had a decreased response to the assay, i.e. a normalized ratio below 0.80. Its specificity was also 100% since none of the other tested patients had a decreased response, i.e. isolated protein C (n = 3) or protein S deficiency (n = 50), or without any abnormality of the protein C. pathway (n = 143), even those on oral anticoagulant treatment (n = 76). However, it would be preferable that each laboratory defines both its reference range and its cutoff level. Finally, even if larger-scale multicentre studies are needed before definite recommendations could be made, these results suggest that the ProC((R)) Global performed using a 1:5 pre-dilution of the patient plasma in factor V-depleted plasma could be validly used as a screening assay of the factor V Leiden mutation-related APC resistance in patients with a history of thrombosis. (C) 2001 Lippincott Williams & Wilkins.