NEW ALLELE-SPECIFIC PRIMERS FOR DETECTING THE LEIDEN MUTATION IN EXON-10 OF THE FACTOR-V GENE IN THROMBOPHILIA

New allele-specific primers were developed which enable the facile and effective identification of the Leiden mutation in the human genome u sing PCR. One of the primers (allele-nonspecific), which is complement ary to the nucleotide sequence of the intron 10 sense strand, [(5')TCT CTTGAAGGAAATGCCCCATTA], was described by B. Dahlback in 1994. Two othe r primers (allele-specific), (5')TAAGAGCAGATCCCTGGACAGCCA and (S')TAAG AGCAGATCCCTGGACACGCA), contained a 3'-terminal nucleotide correspondin g to the nucleotide of the mutant allele, as well as a nucleotide nonc omplementary to the template DNA near the 3'-end (shown by boldface ty pe). When used in combination with allele-nonspecific primers, both al lele-specific primers were equally effective in detecting the Leiden m utation in the human factor V gene. Using these primers, two Leiden mu tations in the heterozygous state were found in 20 patients with deep vein thromboses and pulmonary thromboembolia.