title: [New allele-specific primers for detection of Leiden mutation in exon 10 of factor V gene in thrombophilias].
abstract: New allele-specific primers were developed which enable the facile and effective identification of the Leiden mutation in the human genome using PCR. One of the primers (allele-nonspecific), which is complementary to the nucleotide sequence of the intron 10 sense strand, [(5')TCTCTTGAAGGAAATGCCCCATTA], was described by B. Dahlback in 1994. Two other primers (allele-specific), (5')TAAGAGCAGATCCCTGGACAGCCA and (5')TAAGAGCAGATCCCTGGACACGCA), contained a 3'-terminal nucleotide corresponding to the nucleotide of the mutant allele, as well as a nucleotide noncomplementary to the template DNA near the 3'-end (shown by boldface type). When used in combination with allele-nonspecific primers, both allele-specific primers were equally effective in detecting the Leiden mutation in the human factor V gene. Using these primers, two Leiden mutations in the heterozygous state were found in 20 patients with deep vein thromboses and pulmonary thromboembolia.
authors.forename: E S
authors.initials: ES
authors.lastname: Zykova
authors.forename: L I
authors.initials: LI
authors.lastname: Patrushev
authors.forename: A L
authors.initials: AL
authors.lastname: Kaiushin
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: Bokarev
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; D001778:Blood Coagulation Disorders / Q000097:blood / Q000235:genetics*; D005091:Exons; D005165:Factor V / Q000235:genetics*; D006801:Humans; D009154:Mutation; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction
chemical_list: D015345:Oligonucleotide Probes; C095381:factor V Leiden; D005165:Factor V