title: Genotyping of eight polymorphic genes encoding drug-metabolizing enzymes and transporters using a customized oligonucleotide array.
abstract: Polymorphisms in drug-metabolizing genes may lead to the production of dysfunctional proteins and consequently affect therapeutic efficacy and toxicity of drugs. Different frequencies of polymorphic alleles among the races have been postulated to account for the observed ethnic variations in drug responses. In the current study, we aimed to estimate the frequencies of 14 polymorphisms in eight genes (TPMT, NQO1, MTHFR, GSTP1, CYP1A1, CYP2D6, ABCB1, and SLC19A1) in the Singapore multiracial populations by screening 371 cord blood samples from healthy newborns. To improve genotyping efficacy, we designed an oligonucleotide array based on the principle of allele-specific primer extension (AsPEX) that was capable of detecting the 14 polymorphisms simultaneously. Cross-validation using conventional polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) assays demonstrated 99% concordant results. Measurements on the fluorescent intensity displayed clear distinctions among different genotypes. Statistical analyses showed significantly different allele distributions in several genes among the three races, namely Chinese, Malays, and Indians. Comparing the allelic frequencies in Chinese with previous studies in Caucasian populations, NQO1 609C>T and SLC19A1 80G>A were distinctly different, whereas close similarity was observed for MTHFR 677C>T. We have demonstrated an array-based methodology for rapid multiplex detection of genetic polymorphisms. The allelic frequencies reported in this study may have important therapeutic and prognostic implications in the clinical use of relevant drugs.
authors.affiliation: Department of Pediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119074.
authors.forename: Yi
authors.initials: Y
authors.lastname: Lu
authors.forename: Shirley Kow-Yin
authors.initials: SK
authors.lastname: Kham
authors.forename: Toon-Chai
authors.initials: TC
authors.lastname: Foo
authors.forename: Hany
authors.initials: H
authors.lastname: Ariffin
authors.forename: Ariffin
authors.initials: A
authors.lastname: Hany
authors.forename: Thuan-Chong
authors.initials: TC
authors.lastname: Quah
authors.forename: Allen Eng-Juh
authors.initials: AE
authors.lastname: Yeoh
mesh_terms: D000483:Alleles; D004798:Enzymes / Q000235:genetics*; D005787:Gene Frequency; D005838:Genotype; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D010597:Pharmacogenetics; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D013050:Spectrometry, Fluorescence
chemical_list: D004798:Enzymes