Two multiplex PCR-based DNA assays for the thrombosis risk factors prothrombin G20210A and coagulation factor V G1691A polymorphisms

Two coagulation factor polymorphisms, G1691A in the factor V gene and G2021 0A in the prothrombin gene, are currently the most common known genetic ris k factors for venous thrombosis in caucasian populations. Experimental cond itions that permit simultaneous determination of these two polymorphisms us ing PCR-based DNA assays with the restriction enzyme, Hind III, were develo ped. Moreover, novel PCR-based DNA assays were established using primers th at introduce specific cleavage sequences which then permit the simultaneous determination of these polymorphisms using Sac I restriction digestion ana lysis. Multiplex PCR-based DNA assays were used to analyze a family with a history of venous thrombosis in which these polymorphisms were both present and the results confirm the interesting variability of these genetic risk factors among family members who are symptomatic. (C) 1999 Elsevier Science Ltd. All rights reserved.