Sauer Matthias, Tews Julia, Schwarzer Michael, Seeburger Joerg, Adams Volker, Falk Volkmar, Dhein Stefan, Mohr Friedrich-Wilhelm
Department of Cardiac Surgery, Heart Center, University of Leipzig, Germany.
Clin Lab. 2009;55(3-4):128-36.
The beta1-adrenoceptor plays a crucial role in the regulation of myocardial contractility and heart rate. A functionally important polymorphism at codon 389 has been associated with heart failure and hypertension. The conventional method of detecting this polymorphism is a PCR-assay followed by digestion with specific restriction enzyme (PCR-RFLP) polymorphism. We aimed to establish an alternative, fast genotyping method by real time PCR, which combines high speed DNA-amplification and real time fluorescence monitoring using fluorescence resonance energy transfer. Venous blood of 50 healthy volunteers was examined for the Arg389Gly polymorphism using both methods. The results from PCR-RFLP were compared to those obtained with real time fluorescence PCR and showed a 100% correlation. DNA sequencing confirmed a 100% concordance. This alternative technique is significantly less time consuming than traditional PCR-RFLP and facilitates the analysis of a large numbers of samples.
