title: Mutation in the gene coding for coagulation factor V and resistance to activated protein C: detection of the genetic mutation by oligonucleotide ligation assay using a semi-automated system.
abstract: Resistance of coagulation factor Va to inactivation by activated protein C (APCR) is associated with a point mutation in which adenine is substituted for guanine at nucleotide 1691 in the gene coding for factor V (FV Leiden). To date, this mutation of factor V is the most frequent genetic risk factor for venous thrombophilia. In this report, we describe the adaptation of an automatable oligonucleotide ligation assay (OLA) to detect the mutation in polymerase chain reaction-amplified DNA samples from 40 normal, 20 affected heterozygous, and 3 affected homozygous individuals. The genotypes determined by conventional allele-specific restriction enzyme site analysis were in complete concordance with the results obtained by ELISA-based oligonucleotide-ligation assay. The automated oligonucleotide ligation assay provides a rapid, reliable, nonisotopic method to detect the mutation responsible for APCR that can rapidly be applied to large population screening.
authors.affiliation: Department of Hemostasis and Transfusion Medicine, Heinrich Heine University Medical Center, Düsseldorf, Germany.
authors.forename: R B
authors.initials: RB
authors.lastname: Zotz
authors.forename: B
authors.initials: B
authors.lastname: Maruhn-Debowski
authors.forename: R E
authors.initials: RE
authors.lastname: Scharf
mesh_terms: D004797:Enzyme-Linked Immunosorbent Assay; D005165:Factor V / Q000037:antagonists & inhibitors / Q000235:genetics*; D006801:Humans; D015345:Oligonucleotide Probes; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011486:Protein C / Q000235:genetics* / Q000378:metabolism
chemical_list: D015345:Oligonucleotide Probes; D011486:Protein C; D005165:Factor V