Denaturing high-performance liquid chromatography mutation analysis in patients with reduced Protein S levels

Clin Chim Acta. 2008 Apr;390(1-2):76-81. doi: 10.1016/j.cca.2007.12.023. Epub 2008 Jan 16.

Abstract

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.

Patients: 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.

MeSH terms

  • Base Sequence
  • Chromatography, High Pressure Liquid / methods*
  • DNA Primers
  • Humans
  • Mutation*
  • Polymerase Chain Reaction
  • Protein Denaturation
  • Protein S / metabolism*

Substances

  • DNA Primers
  • Protein S