Rapid detection of the factor XIII Val34Leu (163 G → T) polymorphism by real-time PCR using fluorescence resonance energy transfer detection and melting curve analysis

Amir H. Shemirani, László Muszbek

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21 Citations (Scopus)

Abstract

The Val34Leu polymorphism in the A subunit of blood coagulation factor XIII (FXIII-A) is located in the activation peptide, just three amino acids upstream of the thrombin cleavage site. The Val → Leu replacement accelerates the rate of the proteolytic activation of FXIII and it seems to provide protection against myocardial infarction. Methods available for the assessment of the FXIII-A Val34Leu polymorphism are rather time-consuming, laborious and not easily applicable for large-scale studies. In this study a new method based on real-time PCR with fluorescence resonance energy transfer (FRET) detection and melting curve analysis was developed. The rapid, simple method was adapted to the widely used real-time PCR instrument, LightCycler (Roche Diagnostics). The results showed 100% coincidence with those obtained by the traditional PCR-restriction fragment length polymorphism (RFLP) assay and fluorescent DNA sequencing. Using this method, an allele frequency of 24.2% was obtained (n = 113), which well agrees with the allele frequency obtained by PCR-RFLP on a different group of the same ethnic Hungarian population (25.9%).

Original languageEnglish
Pages (from-to)877-879
Number of pages3
JournalClinical Chemistry and Laboratory Medicine
Volume42
Issue number8
DOIs
Publication statusPublished - Sep 22 2004

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Keywords

  • Factor XIII
  • Fluorescence resonance energy transfer
  • Gene polymorphism
  • Real-time PCR

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

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