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A simple method for the detection of CYP2C9 polymorphisms: nested allele-specific multiplex polymerase chain reaction.

Z Zainuddin·L K Teh·A W M Suhaimi·M Z Salleh·R Ismail
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Abstract

BACKGROUND: Cytochrome P4502C9 (CYP2C9), a principle drug-metabolizing enzyme is polymorphic in humans and is responsible for important pharmacokinetic and pharmacodynamic variations of CYP2C9 substrates. We developed an allele-specific multiplex polymerase chain reaction (PCR) method for the detection of common CYP2C9 alleles. METHOD: Genomic DNA was extracted from blood obtained from 40 unrelated healthy Malaysian Indian volunteers. The DNA was subjected to a first PCR that was used to amplify both exons 3 and 7 simultaneously in one reaction tube and a second PCR that was used to detect the polymorphic sites of CYP2C9 alleles using allele-specific primers. Sequencing was performed to validate the test results. RESULTS: We were successful in amplifying the fragments of interest from the DNA samples. The method was also reproducible and specific. The amplified sequences showed 100% homology to CYP2C9 sequence. CONCLUSION: This is the first nested allele-specific multiplex PCR method reported to allow for the simultaneously detection of five CYP2C9 alleles.

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AllelesAryl Hydrocarbon HydroxylasesCytochrome P-450 CYP2C9DNADNA PrimersElectrophoresis, Agar GelGenotypeHumansMagnesium ChloridePolymerase Chain ReactionPolymorphism, GeneticJournal ArticleResearch Support, Non-U.S. Gov't

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Z Zainuddin, L K Teh, A W M Suhaimi, M Z Salleh, R Ismail. 2003. A simple method for the detection of CYP2C9 polymorphisms: nested allele-specific multiplex polymerase chain reaction.. https://doi.org/10.1016/s0009-8981(03)00319-x

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