title: Automated detection of the factor V Leiden mutation using the LCx microparticle enzyme immunoassay.
abstract: The factor V Leiden mutation, a G-->A transition at position 1691 in exon 10 of the gene that codes for factor V, produces an Arg506Gln substitution and is the most common genetic risk factor for venous thrombosis. We have developed a rapid, sensitive, and specific method to detect the factor V Leiden mutation in genomic DNA from whole blood by PCR amplification and microparticle enzyme immunoassay detection using the Abbott LCx instrument. We compared this automated method with the standard procedure using restriction endonuclease digestion of PCR products followed by gel electrophoresis in blinded experiments. In 130 patients (from Veterans Affairs medical centers) with deep venous thromboses, including 24 heterozygotes with the factor V Leiden mutation, there was complete agreement between the two methods. The assay was also able to distinguish heterozygotes from homozygotes. This method, which carries a low potential for cross-contamination of samples, should be a useful routine test for the factor V Leiden mutation in clinical laboratories with sufficient demand for molecular diagnostic assays using the LCx instrument.
authors.affiliation: Hematology-Oncology Section, Brockton-West Roxbury Veterans Affairs Medical Center, Brockton, MA 02410, USA.
authors.forename: M
authors.initials: M
authors.lastname: Hunault
authors.forename: C
authors.initials: C
authors.lastname: Marsh-Scott
authors.forename: C
authors.initials: C
authors.lastname: Jou
authors.forename: R
authors.initials: R
authors.lastname: Marshall
authors.forename: C
authors.initials: C
authors.lastname: Scheffel
authors.forename: L D
authors.initials: LD
authors.lastname: Fiore
authors.forename: K A
authors.initials: KA
authors.lastname: Bauer
authors.forename: M J
authors.initials: MJ
authors.lastname: Mitchell
mesh_terms: D001322:Autoanalysis; D004252:DNA Mutational Analysis / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005165:Factor V / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008875:Middle Aged; D009154:Mutation; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity
chemical_list: D011933:Reagent Kits, Diagnostic; C095381:factor V Leiden; D005165:Factor V