title: Double fluorescent-amplification refractory mutation detection (dF-ARMS) of the factor V Leiden and prothrombin mutations.
abstract: Simultaneous fluorescent [F] detection of the factor V Leiden (G1691A) and the prothrombin 3'-untranslated region (G20210A) mutations were performed in a single tube polymerase chain reaction (PCR). Amplification refractory mutation detection system (ARMS) formed the basis of this assay design. Fluorescent-labelled primers incorporated into amplicons during the reaction facilitated detection directly by GeneScan analysis without further manipulation. To test the efficacy of this double [F]-ARMS (dF-ARMS) method, 48 patients with unexplained thrombotic tendencies were investigated for their factor V Leiden and prothrombin genotypes. These results corresponded exactly with data achieved using the more conventional methods of restriction fragment length polymorphism (RFLP)-PCR and direct DNA sequencing. Three out of the 48 patients in this group were found to be compound heterozygotes.
authors.affiliation: Molecular Diagnostics Unit, Cork Institute of Technology, Ireland.
authors.forename: C
authors.initials: C
authors.lastname: Maher
authors.forename: D
authors.initials: D
authors.lastname: Crowley
authors.forename: C
authors.initials: C
authors.lastname: Cullen
authors.forename: C
authors.initials: C
authors.lastname: Wall
authors.forename: D
authors.initials: D
authors.lastname: Royston
authors.forename: S
authors.initials: S
authors.lastname: Fanning
mesh_terms: D000483:Alleles; D005165:Factor V / Q000235:genetics*; D005453:Fluorescence; D006801:Humans; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011516:Prothrombin / Q000235:genetics*