Duplex PCR-RFLP for simultaneous detection of factor V Leiden and prothrombin G20210A.

The authors have used a primer-engineered duplex polymerase chain reaction (PCR) -restriction fragment length polymorphism (RFLP) for the simultaneous detection of factor V Leiden and prothrombin G20210A mutations. The method involves the generation of HindIII RFLPs, and the restricted amplification products are analyzed by agarose gel electrophoresis in a single gel lane. This method is simple an inexpensive, and readily adaptable to the routine in a clinical molecular diagnostic laboratory other that our own.