Denaturing high-performance liquid chromatography mutation analysis in patients with reduced Protein S levels.
BACKGROUND
Patients with congenital Protein S deficiency have increased risk of venous thromboembolism. However, Protein S levels show large intra-individual variation and the biochemical assays have low accuracy and a high interlaboratory variability. Genetic analysis might aid in a more precise diagnosis and risk estimation. The aim was to design a high-throughput genetic analysis based on denaturing high-performance liquid chromatography to identify sequence variations in the gene coding for Protein S.


METHODS
In total, 55 patients referred to the Section of Thrombosis and Haemostasis, Odense University Hospital, in the period 1998-2004 were included in the study.


RESULTS
Mutations were found in ten of the 55 patients: Six different variants were identified, of which four were not previously reported: One were a nonsense mutation substituting a glutamine with a stopcodon (c.790C>T) and the rest were missense mutations (c.932T>G; c.1367A>G; c.1378T>C). Furthermore, four patients carried the same mutation (c.1045G>A), while two carried the Heerlen mutation (c.1378T>C).


CONCLUSIONS
The reported method will be useful for rapidly detecting sequence variations in the gene coding for Protein S, giving a precise diagnosis and subsequently a better risk estimation.
Bathum
Lise
L
Department of Biochemistry, Pharmacology and Genetics, Odense University Hospital, Denmark. LisBat@vestamt.dk
Münster
Anna-Marie
AM
Nybo
Mads
M
Madsen
Jonna Skov
JS
Larsen
Torben Bjerregaard
TB
Kristensen
Søren Risom
SR
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017931:DNA Primers; D006801:Humans; D009154:Mutation; D016133:Polymerase Chain Reaction; D011489:Protein Denaturation; D017293:Protein S / Q000378:metabolism*
D017931:DNA Primers; D017293:Protein S