Objectives: Numerous functional polymorphisms in the CYP2C19 gene have been identified; some alleles (e.g. CYP2C19*2 and CYP2C19*3) are associated with poor metabolism of CYP2C19 substrate drugs. Studies have found that the proportion of poor metabolizers, explained by CYP2C19*2 and CYP2C19*3, varies from less than 50% to more than 90% of poor metabolizers. Therefore, phenotype-genotype correlation studies should cover more than CYP2C19*2 and CYP2C19*3. A broader coverage, however, requires an easy-to-use and high-throughput genotyping platform. This broader coverage should also include the recently identified functional allele, CYP2C19*10, which involves a nucleotide change adjacent to the altered nucleotide change in CYP2C19*2. The currently used restriction fragment length polymorphism-based method for genotyping CYP2C19*2 cannot distinguish between CYP2C19*2 and CYP2C19*10. We aim to develop a simple platform that can genotype all CYP2C19 functional alleles.
Methods: We have developed a thin-film biosensor chip platform to genotype 16 exonic CYP2C19 variants, including two sets of two adjacent single nucleotide polymorphisms and 12 single single nucleotide polymorphisms, using a ligation strategy.
Results: We demonstrate that this is a rapid, accurate, and inexpensive method for genotyping CYP2C19 variants using individual's genomic DNA samples. We further demonstrate that this genotyping platform can be used to construct a haplotype structure of the CYP2C19 variants in a population, and to assign a haplotype combination to each individual on the basis of his/her genotype results.
Conclusion: This assay can be applied in pharmacogenomic studies in both basic research and clinical laboratories. It is also an ideal technology for pharmacogenomic tests in both developed and developing countries.