Factor V Leiden mutation screened by PCR and detected with lanthanide-labeled probes

Genet Test. 2001 Winter;5(4):291-7. doi: 10.1089/109065701753617417.

Abstract

The Factor V Leiden mutation is an important human polymorphism, responsible for increased risk of venous thrombosis in heterozygotes as well as homozygotes. Therefore, screening is a useful possibility, and many detection systems have been described for PCR products. We have developed a simplified and robust assay using oligonucleotide probes for normal and mutant sequences, labeled with europium and samarium, respectively, and measured by time-resolved fluorescence. Populations consisting of 233 Welsh and 148 Irish subjects were examined by both restriction fragment length polymorphism (RFLP) analysis and our assay. The allele frequency was 14/466 in the Welsh and 5/296 in the Irish population, in line with other surveys of European populations. Results were not obtained in 2/381 samples by RFLP, compared with 1/381 with our method. We conclude that our method represents an improved system capable of considerable throughput at reasonable cost.

MeSH terms

  • Factor V / genetics*
  • Genetic Testing / methods
  • Humans
  • Lanthanoid Series Elements*
  • Molecular Probes*
  • Mutation*
  • Polymerase Chain Reaction
  • Staining and Labeling
  • United Kingdom

Substances

  • Lanthanoid Series Elements
  • Molecular Probes
  • factor V Leiden
  • Factor V