[New allele-specific primers for detection of Leiden mutation in exon 10 of factor V gene in thrombophilias].
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.
Zykova
E S
ES
Patrushev
L I
LI
Kaiushin
A L
AL
Korosteleva
M D
MD
Miroshnikov
A I
AI
Bokarev
I N
IN
Leont'ev
S G
SG
Koshkin
V M
VM
Severin
E S
ES
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
D015345:Oligonucleotide Probes; C095381:factor V Leiden; D005165:Factor V