A genotyping method for VKORC1 1173C > T by Pyrosequencing technology

Scand J Clin Lab Invest. 2008;68(5):427-30. doi: 10.1080/00365510701810621.

Abstract

Vitamin K epoxide reductase complex subunit 1 (VKORC1) is the site of inhibition by warfarin and other antivitamin K drugs during oral anticoagulant therapy. The SNP rs9934438 in intron 1 of VKORC1 (c.173+1000C > T or 1173C > T) discriminating the VKORC1*2 haplotype is associated with low warfarin dose requirement and unstable prothrombin time - international normalized ratio. To genotype this SNP, we have developed a rapid method using Pyrosequencing technology. The proposed method takes a post-PCR sample preparation of less than 1 h and a DNA sequencing time of less than 15 min to genotype 96 samples. The current method was compared with a dHPLC method that we reported previously. Genotype frequencies at VKORC1 1173C>T for our Swedish population were 38% wild-type, 40% heterozygote and 22% homozygote. The frequency of the T-allele was 0.42, which exactly matches the frequency previously reported for Germans. The current method can be used to determine whether patients initiating warfarin therapy are carriers of SNP 1173 C>T that is strongly associated with low warfarin dose requirement.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Mutational Analysis / methods*
  • Genotype
  • Humans
  • Mixed Function Oxygenases / genetics*
  • Polymorphism, Single Nucleotide / genetics*
  • Vitamin K Epoxide Reductases

Substances

  • Mixed Function Oxygenases
  • VKORC1 protein, human
  • Vitamin K Epoxide Reductases