Genotyping and mapping assay of single-nucleotide polymorphisms in CYP3A5 using DNA-binding zinc(II) complexes.

Objective: It is clinically important to detect the single-nucleotide polymorphism (SNP) of CYP3A53 (6986A>G) associated with enzymatic activity for drug metabolism. The aim of this study was to establish an accurate strategy for SNP screening.

Design And Methods: Polyacrylamide gel electrophoresis (PAGE) using zinc(II) complexes were applied for SNP detection. Genomic analyses of 19 healthy subjects were conducted by using both Zn(2+)-Phos-tag-PAGE and Zn(2+)-cyclen-PAGE methodologies.

Results: Zn(2+)-Phos-tag PAGE permitted identification of the following allele genotypes: the A/A homozygote (CYP3A51/1) in 3 individuals, the G/G homozygote (CYP3A53/3) in 14 individuals, and the A/G heterozygote (CYP3A51/3) in 2 individuals. Zn(2+)-cyclen PAGE demonstrated not only reproducibility of the genotyping but also existence of a novel heterozygous SNP (6929G>A) in the subject with CYP3A51/1.

Conclusion: We demonstrated reliable SNP genotyping and mapping in CYP3A5 using the combination method of Zn(2+)-Phos-tag PAGE and Zn(2+)-cyclen PAGE.