title: Factor V Leiden mutation screened by PCR and detected with lanthanide-labeled probes.
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.
authors.affiliation: BioQuant Ltd, Oxford BioBusiness Centre, Littlemore, Oxford, OX4 4SS, UK. colin@bioquant.co.uk
authors.forename: C G
authors.initials: CG
authors.lastname: Potter
authors.forename: Y T
authors.initials: YT
authors.lastname: Liu
authors.forename: D C
authors.initials: DC
authors.lastname: Rees
mesh_terms: D005165:Factor V / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D028581:Lanthanoid Series Elements; D015335:Molecular Probes; D009154:Mutation; D016133:Polymerase Chain Reaction; D013194:Staining and Labeling; D006113:United Kingdom
chemical_list: D028581:Lanthanoid Series Elements; D015335:Molecular Probes; C095381:factor V Leiden; D005165:Factor V