An Allele-Specific PCR System for Rapid Detection and Discrimination of the CYP2C19*4A, *4B, and*17 Alleles Implications for Clopidogrel Response Testing

作者：Scott Stuart A^*; Tan Qian; Baber Usman; Yang Yao; Martis Suparna; Bander Jeffrey; Kornreich Ruth; Hulot Jean Sebastien; Desnick Robert J

来源：Journal of Molecular Diagnostics, 2013, 15(6): 783-789.

DOI：10.1016/j.jmoldx.2013.06.004

CYP2C19 is involved in the metabolism of clinically relevant drugs, including the antiplatelet prodrug clopidogrel, which has prompted interest in clinical CYP2C19 genotyping. The CYP2C19*48 allele is defined by both gain-of-function [c.-806C%26gt;T (*17)] and Loss-of-function [c.1A%26gt;G (*4)] variants on the same haptotype; however, current genotyping and sequencing assays are unable to determine the phase of these variants. Thus, the aim of this study was to develop an assay that could rapidly detect and discriminate the related *4,4, *4B, and *17 alleles. An allele-specific PCR assay, composed of four unique primer mixes that specifically interrogate the defining *17 and *4 variants, was developed by using samples (n = 20) with known genotypes, including the *4A, *48, and/or *17 alleles. The assay was validated by testing 135 blinded samples, and the results were correlated with CYP2C19 genotyping and allele-specific cloning/sequencing. Importantly, among the six *4 carriers in the validation cohort, after allele-specific PCR testing both samples with a *14 genotype were reclassified to *17*4A, all three samples with a *417 genotype were reclassified to *148, and a sample with a *41717 genotype was reclassified to *4817. In conclusion, this rapid and robust allele-specific PCR assay can refine CYP2C19 genotyping and metabolizer phenotype classification by determining the phase of the defining *17 and *4 variants, which may have utility when testing CYP2C19 for ctopidogrel response.

• 出版日期2013-11