An improved rapid genotyping method for the study of beta-2 adrenergic receptor gene polymorphisms using single tube allele specific multiplex PCR.
BACKGROUND
Seventeen single nucleotide polymorphisms (SNPs) have been identified so far, within the beta-2 receptor (beta(2) AR) gene. The presence of so many SNPs within the beta(2) AR gene causes a problem, for those studying beta(2) AR pharmacogenetics, in relation to which SNPs to choose. Most of the work has focused on the three common SNPs within the coding block (alleles 16, 27 and 164) and the techniques developed have been for these three functionally important alleles.


OBJECTIVE
We report an improved polymerase chain reaction (PCR)-based method for the simultaneous detection of five functionally important beta(2) AR alleles, namely beta 16A/G, beta utr-20C/T, beta 27C/G, beta utr-47C/T and beta 164C/T.


METHODS
Genomic DNA was used as a template for duplex and triplex PCR to detect the polymorphic sites of the five alleles.


RESULTS
DNA sequencing analysis confirmed the specificity of this PCR method.


CONCLUSIONS
This simplified single-tube multiplexed PCR assay provides an easier, faster and more cost-effective method than those available for studying the specified polymorphisms of the beta(2)AR gene.
Zilfalil
B A
BA
Human Genome Centre, School of Medical Science, Universiti Sains Malaysia, Kota Bharu, Malaysia. zilfalil@kb.usm.my
Hoh
B P
BP
Nizam
M Z
MZ
Liza-Sharmini
A T
AT
Teh
L K
LK
Ismail
R
R
D005838:Genotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics*
D018343:Receptors, Adrenergic, beta-2