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%).
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Authors | Amir H Shemirani, László Muszbek |
Journal | Clinical chemistry and laboratory medicine
(Clin Chem Lab Med)
Vol. 42
Issue 8
Pg. 877-9
( 2004)
ISSN: 1434-6621 [Print] Germany |
PMID | 15387436
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
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Topics |
- Alleles
- DNA
(chemistry, genetics)
- Factor XIII
(analysis, chemistry, genetics)
- Fluorescence Resonance Energy Transfer
(methods)
- Gene Frequency
- Humans
- Hungary
- Nucleic Acid Denaturation
- Polymerase Chain Reaction
(instrumentation, methods)
- Polymorphism, Genetic
- Polymorphism, Restriction Fragment Length
- Temperature
- Time Factors
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