Rapid genotyping of haemostatic gene polymorphisms using the 5' nuclease assay.
Hemostatic gene polymorphisms have been shown to be associated with arterial and venous thrombotic disease. To date these polymorphisms have mainly been detected by labor-intensive conventional gel based methods. Aim of the present study was to design and optimize high throughput 5' nuclease assays for the detection of a set of 10 single-nucleotide polymorphisms (SNP) in genes of importance for hemostasis: plasminogen activator inhibitor type 1 -675 4G>5G, thrombin activatable fibrinolysis inhibitor Ala147Thr and 1,542C>G, beta-fibrinogen -455G>A, von Willebrand factor -1,051A>G, factor VII Arg353Gln, factor XIII Val34Leu, prothrombin 20,210G>A, tissue factor pathway inhibitor -287T>C, and methylenetetrahydrofolate reductase 1,298A>C. Specificity of each genotyping assay was confirmed by sequence-based typing and reproducibility was evaluated by repeated genotyping. The genotyping protocols presented here may serve as a valuable tool for clinical researchers interested in exploring associations between these SNPs and thrombotic disease.
Department of Clinical Genetics, Sahlgrenska University Hospital, Sweden. christina.jern@neuro.gu.se
Anna
A
Tjärnlund
Maria
M
Andersson
Katarina
K
Jood
Per
P
Ladenvall
Christina
C
Jern
D001779:Blood Coagulation Factors / Q000235:genetics; D001798:Blood Proteins / Q000235:genetics; D003936:Diagnosis, Computer-Assisted; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006487:Hemostasis / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D019851:Thrombophilia / Q000175:diagnosis / Q000235:genetics
D001779:Blood Coagulation Factors; D001798:Blood Proteins