SNP genotyping in the β2-adrenergic receptor by electronic microchip assay, DHPLC, and direct sequencing

Abstract

The β2-adrenergic receptor (β2AR) is the key target for the β2-agonist drugs used for bronchodilation in asthma and chronic obstructive pulmonary disease. To detect four SNPs with amino acid variations at positions -47T/C (CysBUP19Arg), 46A/G (Gly16Arg), 79C/G (Gln27Glu), and 491C/T (Thr164Ile) in the β2AR gene, we used the electronic microchip assay, denaturing high-performance liquid chromatography (DHPLC), and direct sequencing. Genomic DNA samples were obtained from the blood of 84 Japanese healthy volunteers. The agreement rates of the first data set with the final data (allele calls) were 99.7% (332/333), 99.2% (246/248), and 96.7% (329/340). The percentages of no allele designation (ND) were 2.06% (7/340), 2.75% (7/255), and 0.00% (0/340) for the electronic microchip assay, DHPLC, and direct sequencing, respectively. Furthermore, we found three samples that had a novel haplotype.