title: Use of first nucleotide change technology to determine the frequency of factor V Leiden in a population of Australian blood donors.
abstract: Activated protein C resistance (APCR), the most common risk factor for venous thrombosis, is the result of a G to A base substitution at nucleotide 1691 (R506Q) in the factor V gene. Current techniques to detect the factor V Leiden mutation, such as determination of restriction length polymorphisms, do not have the capacity to screen large numbers of samples in a rapid, cost-effective test. The aim of this study was to apply the first nucleotide change (FNC) technology, to the detection of the factor V Leiden mutation. After preliminary amplification of genomic DNA by polymerase chain reaction (PCR), an allele-specific primer was hybridised to the PCR product and extended using fluorescent terminating dideoxynucleotides which were detected by colorimetric assay. Using this ELISA-based assay, the prevalence of the factor V Leiden mutation was determined in an Australian blood donor population (n = 500). A total of 18 heterozygotes were identified (3.6%) and all of these were confirmed with conventional MnlI restriction digest. No homozygotes for the variant allele were detected. We conclude from this study that the frequency of 3.6% is compatible with others published for Caucasian populations. In addition, the FNC technology shows promise as the basis for a rapid, automated DNA based test for factor V Leiden.
authors.affiliation: Co-operative Research Centre for Diagnostic Technologies, School of Life Sciences, Queensland University of Technology, Brisbane, Australia.
authors.forename: N M
authors.initials: NM
authors.lastname: Pecheniuk
authors.forename: N A
authors.initials: NA
authors.lastname: Marsh
authors.forename: T P
authors.initials: TP
authors.lastname: Walsh
authors.forename: J L
authors.initials: JL
authors.lastname: Dale
mesh_terms: D000225:Adenine / Q000032:analysis*; D001315:Australia / Q000453:epidemiology; D001782:Blood Donors; D005165:Factor V / Q000378:metabolism*; D005820:Genetic Testing / Q000379:methods*; D014644:Genetic Variation; D006147:Guanine / Q000032:analysis*; D006801:Humans; D009154:Mutation; D015995:Prevalence; D012307:Risk Factors; D019851:Thrombophilia / Q000097:blood* / Q000453:epidemiology
chemical_list: C095381:factor V Leiden; D006147:Guanine; D005165:Factor V; D000225:Adenine