Real-time PCR for rapid genotyping of angiotensin-converting enzyme insertion/deletion polymorphism.
OBJECTIVE
To develop a real-time PCR technique for detection of the insertion/deletion (I/D) polymorphism of angiotensin-converting enzyme (ACE) gene.


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 73.9 degrees C and 76.2 degrees C 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.
Lin
M H
MH
School of Medical Technology, Chang Gung University, Taiwan, Republic of China.
Tseng
C H
CH
Tseng
C C
CC
Huang
C H
CH
Chong
C K
CK
Tseng
C P
CP
D000483:Alleles; D017353:Gene Deletion; D005838:Genotype; D006801:Humans; D016254:Mutagenesis, Insertional; D007703:Peptidyl-Dipeptidase A / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic
D007703:Peptidyl-Dipeptidase A