A multiplex assay for detecting Genetic variations in cyp2c9, vkorc1, and GGCX involved in warfarin metabolism Journal Article
| Authors: | Rai, A. J.; Udar, N.; Saad, R.; Fleisher, M. |
| Article Title: | A multiplex assay for detecting Genetic variations in cyp2c9, vkorc1, and GGCX involved in warfarin metabolism |
| Abstract: | BACKGROUND: Patients differ in responses to warfarin, which is commonly prescribed to treat thromboembolic events. Genetic variations in the cytochrome P450, family 2, subfamily C, polypeptide 9 (CYP2C9), vitamin K epoxide reductase complex, subunit 1 (VKORC1), and gamma-glutamyl carboxylase (GGCX) genes have been shown to contribute to impaired metabolism of warfarin. METHODS: We designed a custom multiplex singlenucleotide polymorphism (SNP) panel to interrogate the CYP2C9 2, 3, VKORC1 (-1639G→A), and GGCX (1181T→G) alleles simultaneously in a single sample by use of single-base extension and capillary electrophoresis after genomic DNA extraction and PCR amplification. RESULTS: Our assay successfully detected various genotypes from known controls and 24 unknown samples. It was found to be 100% concordant with sequencing results. CONCLUSIONS: Our multiplexed SNP panel can be successfully used in genotyping of patient blood samples. Results can be combined with other clinical parameters inan algorithm for warfarin dosing. These data provide a proof-in-principle of multiplexed SNP analysis using rapid single-base extension and capillary electrophoresis, and warrant additional validation using a larger cohort of patient samples. |
| Keywords: | controlled study; unclassified drug; single nucleotide polymorphism; genetics; polymorphism, single nucleotide; polymerase chain reaction; metabolism; allele; gene; gene amplification; genetic variability; genotype; blood sampling; quantitative analysis; gene identification; capillary electrophoresis; warfarin; dna sequence; cytochrome p450 2c9; drug metabolism; aryl hydrocarbon hydroxylases; genetic techniques; genetic procedures; dna extraction; genomic dna; carboxylic acid derivative; gamma glutamyl carboxylase; menadione epoxide; vitamin k epoxide reductase complex subunit 1; cyp2c9 protein, human; glutamyl carboxylase; ligase; menadione epoxidase; mixed function oxidase; unspecific monooxygenase; carbon-carbon ligases; mixed function oxygenases |
| Journal Title: | Clinical Chemistry |
| Volume: | 55 |
| Issue: | 4 |
| ISSN: | 0009-9147 |
| Publisher: | American Association for Clinical Chemistry |
| Date Published: | 2009-04-01 |
| Start Page: | 823 |
| End Page: | 826 |
| Language: | English |
| DOI: | 10.1373/clinchem.2008.118497 |
| PUBMED: | 19233910 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 1" - "Export Date: 30 November 2010" - "CODEN: CLCHA" - "Source: Scopus" |
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