Simultaneous determination of methylenetetrahydrofolate reductase C677T and factor V G1691A genotypes by mutagenically separated PCR and multiple-injection capillary electrophoresis

Clin Chem. 1998 Feb;44(2):264-9.

Abstract

The C677T mutation of the methylenetetrahydrofolate reductase gene and the G1691A (Leiden) mutation of the factor V gene are established risk factors for thromboembolic disease. We here present an assay for the simultaneous genotyping of these common genetic variants. The assay involves a strategy based on multiplex mutagenically separated PCR performed in a single tube containing six primers. Separation of the resulting four PCR products (197, 207, 233, and 246 bp) was performed by capillary electrophoresis coupled with laser-induced fluorescence detection. 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Automation
  • DNA Mutational Analysis / methods*
  • DNA Primers / chemistry
  • Electrophoresis, Capillary
  • Factor V / genetics*
  • Fluorescence
  • Genotype
  • Homocysteine / blood
  • Lasers
  • Methylenetetrahydrofolate Reductase (NADPH2)
  • Oxidoreductases Acting on CH-NH Group Donors / genetics*
  • Polymerase Chain Reaction
  • Risk Factors
  • Thrombosis / genetics

Substances

  • DNA Primers
  • factor V Leiden
  • Homocysteine
  • Factor V
  • Oxidoreductases Acting on CH-NH Group Donors
  • Methylenetetrahydrofolate Reductase (NADPH2)