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Cross-linking hybridization assay for direct detection of factor V Leiden mutation 29th Annual Oak Ridge Conference Zehnder, J., VANATTA, R., Jones, C., Sussman, H., Wood, M. AMER ASSOC CLINICAL CHEMISTRY. 1997: 1703–8

Abstract

A nucleic acid photocross-linking technology was used in the development of a direct assay for factor V Leiden, a point mutation in the factor V gene (G1691A) that is the most common inherited risk factor for thrombosis. This cross-linking hybridization assay included two allele-specific capture probes and six signal-generating reporter probes; all were modified with a photoactivated cross-linking compound. By using two different capture probes complementary to a 16-base sequence at the factor V Leiden mutation site, but differing in the nucleotide opposite the mutation site (C vs T), wild-type and factor V Leiden alleles were differentiated in purified DNA specimens. The assay was also successfully applied to genomic DNA in leukocytes isolated from whole blood; the factor V status of 122 patients as determined by this method was in complete concordance with a standard PCR-based assay and clearly discriminated between healthy individuals and factor V Leiden heterozygotes.

View details for Web of Science ID A1997XV89700033

View details for PubMedID 9299963