SIMULTANEOUS-DETERMINATION OF METHYLENETETRAHYDROFOLATE REDUCTASE C677T AND FACTOR-V G1691A GENOTYPES BY MUTAGENICALLY SEPARATED PCR AND MULTIPLE-INJECTION CAPILLARY ELECTROPHORESIS

The C677T mutation of the methylenetetrahydrofolate reductase gene and the G1691A (Leiden) mutation of the factor V gene are established ris k factors for thromboembolic disease. We here present an assay for the simultaneous genotyping of these common genetic variants. The assay i nvolves a strategy based an multiplex mutagenically separated PCR perf ormed in a single tube containing six primers. Separation of the resul ting four PCR products (197, 207, 233, and 246 bp) was performed by ca pillary electrophoresis coupled with laser-induced fluorescence detect ion. The time for the automated electrophoresis was reduced to 2.5 min per sample by performing the capillary electrophoresis analysis in a multiple-injection mode.