Rapid detection of the R389G polymorphism of ADRB1 by real time PCR using fluorescence resonance energy transfer.
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.
Sauer
Matthias
M
Department of Cardiac Surgery, Heart Center, University of Leipzig, Germany.
Tews
Julia
J
Schwarzer
Michael
M
Seeburger
Joerg
J
Adams
Volker
V
Falk
Volkmar
V
Dhein
Stefan
S
Mohr
Friedrich-Wilhelm
FW
D000328:Adult; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005787:Gene Frequency; D005838:Genotype; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D018342:Receptors, Adrenergic, beta-1 / Q000235:genetics*; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA
D018342:Receptors, Adrenergic, beta-1