MUTATION IN THE GENE CODING FOR COAGULATION-FACTOR V AND RESISTANCE TO ACTIVATED PROTEIN-C - DETECTION OF THE GENETIC MUTATION BY OLIGONUCLEOTIDE LIGATION ASSAY USING A SEMIAUTOMATED SYSTEM

Resistance of coagulation factor Va to inactivation by activated prote in C (APCR) is associated with a point mutation in which adenine is su bstituted for guanine at nucleotide 1691 in the gene coding for factor V (FV Leiden). To date, this mutation of factor V is the most frequen t genetic risk factor for venous thrombophilia. In this report, we des cribe the adaptation of an automatable oligonucleotide ligation as say (OLA) to detect the mutation in polymerase chain reaction-amplified D NA samples from 40 normal, 20 affected heterozygous, and 3 affected ho mozygous individuals. The genotypes determined by conventional allele- specific restriction enzyme site analysis were in complete concordance with the results obtained by ELISA-based oligonucleotide-ligation ass ay. The automated oligonucleotide ligation assay provides a rapid, rel iable, nonisotopic method to detect the mutation responsible for APCR that can rapidly be applied to large population screening.