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


Sauer, M; Tews, J; Schwarzer, M; Seeburger, J; Adams, V; Falk, V; Dhein, S; Mohr, F W (2009). Rapid detection of the R389G polymorphism of ADRB1 by real time PCR using fluorescence resonance energy transfer. Clinical Laborator, 55(3-4):128-136.

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.

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.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > University Hospital Zurich > Clinic for Cardiovascular Surgery
Dewey Decimal Classification:610 Medicine & health
Language:English
Date:2009
Deposited On:02 Dec 2009 11:55
Last Modified:05 Apr 2016 13:33
Publisher:Verlag Klinisches Labor
ISSN:1433-6510
Related URLs:http://www.clin-lab-publications.com/abstracts/348.html (Publisher)
PubMed ID:19462935

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