Rapid detection of the R389G polymorphism of ADRB1 by real time PCR using fluorescence resonance energy transfer

Clin Lab. 2009;55(3-4):128-36.

Abstract

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.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Female
  • Fluorescence Resonance Energy Transfer / methods*
  • Gene Frequency
  • Genotype
  • Humans
  • Male
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Genetic
  • Polymorphism, Restriction Fragment Length
  • Receptors, Adrenergic, beta-1 / genetics*
  • Reproducibility of Results
  • Sequence Analysis, DNA

Substances

  • Receptors, Adrenergic, beta-1