Genetic determinants of heritable venous thrombosis: genotyping methods for factor V(Leiden)A1691G, methylenetetrahydrofolate reductase C677T, prothrombin G20210A mutation, and algorithms for venous thrombosis investigations.
OBJECTIVE
To implement cost effective and clinically relevant thrombophilic genotyping and homocysteine analysis in our coagulation laboratory.


METHODS
We describe genotyping assays for three of the genetic defects associated with hereditary thrombosis: factor V(Leiden) A1691G, methylenetetrahydrofolate reductase C677T, and prothrombin gene G20210A. A second confirmatory assay for factor V(Leiden) using allele specific oligonucleotide polymerase chain reaction is also presented. We suggest an algorithm for the rational integration of the traditional assays routinely used to investigate venous thrombosis with genotyping and plasma homocysteine measurements.


RESULTS
These polymerase chain reaction based assays were designed to be performed under identical reaction conditions, permitting simultaneous setup, amplification, digestion, and analysis.


CONCLUSIONS
The three genotyping assays presented are robust and relatively easy to perform. Use of an algorithm will ensure efficient resource utilization and minimize unnecessary testing.
Donnelly
J G
JG
Department of Laboratory Medicine, Ottawa Hospital, Faculty of Medicine, University of Ottawa, Ontario, Canada. jdonnelly@civich.ottawa.on.ca
Rock
G A
GA
D045103:5,10-Methylenetetrahydrofolate Reductase (FADH2); D000465:Algorithms; D001483:Base Sequence; D017931:DNA Primers; D005165:Factor V / Q000235:genetics*; D020022:Genetic Predisposition to Disease; D006710:Homocysteine / Q000097:blood; D006801:Humans; D042965:Methylenetetrahydrofolate Reductase (NADPH2); D009154:Mutation; D010088:Oxidoreductases / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011516:Prothrombin / Q000235:genetics*; D020246:Venous Thrombosis / Q000235:genetics*
D017931:DNA Primers; C095381:factor V Leiden; D006710:Homocysteine; D005165:Factor V; D011516:Prothrombin; D010088:Oxidoreductases; D045103:5,10-Methylenetetrahydrofolate Reductase (FADH2); D042965:Methylenetetrahydrofolate Reductase (NADPH2)