Real-Time Pcr for Rapid Genotyping of Angiotensin-Converting Enzyme Insertion/Deletion Polymorphism
Resource
CLINICAL BIOCHEMISTRY v.34 n.8 pp.661-666
Journal
CLINICAL BIOCHEMISTRY
Journal Volume
v.34
Journal Issue
n.8
Pages
661-666
Date Issued
2001
Date
2001
Author(s)
TSENG, CHIN-HSIAO
Abstract
Objective: To develop a real-time PCR technique for detection of the insertion/deletion (YD) polymorphism of angiotensin-converting enzyme (ACE ) gene.Design and methods: Three primers were designed for performing real -time PCR in the presence of SYBR Green I as flurochrome followed by melting curve analysis. Forty human genomic DNA that have been genotyped by two-rounds of conventional PCR were used for evaluation of this technique.Results: Melting curve analysis indicated the melting peak at 73 .9degreesC and 76.2 degreesC corresponding to the presence of I and D alleles, respectively. Comparable genotyping results were obtained by both conventional and real-time PCR. Besides, the mistyping of ID allele individuals by the first run of conventional PCR were accurately genotyped by single-tube real time PCR. Conclusions: The real-time PCR method presented in this study provides a rapid and sensitive way for genotyping of ACE gene that may be suitable for large-scale clinical and epidemiologic study. (C) 2002 The Canadian Society of Clinical Chemists. All rights reserved.
Subjects
angiotensin-converting enzyme
real-time PCR
insertion/ deletion polymorphism
DELETION POLYMORPHISM
HEART-DISEASE
ACE GENE
SDGs
Type
journal article