Thromb Haemost 1999; 82(04): 1294-1296
DOI: 10.1055/s-0037-1614378
Review Article
Schattauer GmbH

Allelic Discrimination of Factor V Leiden Using a 5’ Nuclease Assay

D. Happich
1   From the Institute of Experimental Haematology and Transfusion Medicine, University of Bonn, Germany
,
R. Schwaab
1   From the Institute of Experimental Haematology and Transfusion Medicine, University of Bonn, Germany
,
P. Hanfland
1   From the Institute of Experimental Haematology and Transfusion Medicine, University of Bonn, Germany
,
D. Hoernschemeyer
1   From the Institute of Experimental Haematology and Transfusion Medicine, University of Bonn, Germany
› Institutsangaben
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Publikationsverlauf

Received 20. Februar 1999

Accepted after revision 21. Mai 1999

Publikationsdatum:
08. Dezember 2017 (online)

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Summary

The G1691A (Leiden) mutation of the factor V gene is the most prevalent identified cause of venous thrombosis. Therefore, we developed a new genetic test using the TaqMan system. With this assay which combines PCR amplification and detection reaction in one closed tube, a cohort of 234 patients with a history of thrombosis was screened. In parallel, amplification products of the same patients were screened with a previously described test using endonuclease digestion of PCR products followed by gel electrophoresis. Identical results were obtained by both methods. Among cases, 122 (52%) individuals were homozygous normal, 99 (42%) were heterozygous affected and 13 (5.5%) showed homozygous pattern for the Factor V Leiden mutation. Thus, it could be demonstrated that the new TaqMan assay is a robust, rapid and automated method for high throughput application which avoids time consuming and difficult post-PCR steps.