title: New DNA diagnostic system for detection of factor V Leiden.
abstract: Use was made of allele-specific PCR to develop a highly effective DNA diagnostic system for detection of the factor V Leiden mutation in exon 10 of human factor V gene. The allele-specific primers contain a 3'-OH end nucleotide, which matches a mutant or wild-type nucleotide of the template DNA (A-allele and G-allele, respectively) and also one mismatched nucleotide near the 3'-end. The universal primers have an internal mismatch with the mutant nucleotide of the template DNA and another mismatched nucleotide at 3'-OH end. The A-allele-specific primer enables the preferential amplification of both the homozygous and heterozygous mutant alleles. The extension of the G-allele-specific primer or the universal one is inhibited in the presence of the homozygous factor V Leiden. The developed assay system allowed us to detect five patients, who are heterozygous for factor V Leiden among the 48 patients with deep venous thrombosis and pulmonary thromboembolism.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow. patrush@ibch.siobc.ras.ru
authors.forename: L I
authors.initials: LI
authors.lastname: Patrushev
authors.forename: E S
authors.initials: ES
authors.lastname: Zykova
authors.forename: A L
authors.initials: AL
authors.lastname: Kayushin
authors.forename: M D
authors.initials: MD
authors.lastname: Korosteleva
authors.forename: A I
authors.initials: AI
authors.lastname: Miroshnikov
authors.forename: I N
authors.initials: IN
authors.lastname: Bokarew
authors.forename: S G
authors.initials: SG
authors.lastname: Leont'ev
authors.forename: V M
authors.initials: VM
authors.lastname: Koshkin
authors.forename: E S
authors.initials: ES
authors.lastname: Severin
mesh_terms: D000483:Alleles; D001681:Biological Assay / Q000379:methods*; D004247:DNA / Q000032:analysis*; D005165:Factor V / Q000032:analysis* / Q000235:genetics*; D006801:Humans; D015345:Oligonucleotide Probes; D017354:Point Mutation
chemical_list: D015345:Oligonucleotide Probes; C095381:factor V Leiden; D005165:Factor V; D004247:DNA