Rapid single-nucleotide Polymorphism detection of cytochrome P450 (CYP2C9) and vitamin k epoxide reductase (VKORC1) genes for the warfarin dose adjustment by the Smart-Amplification Process Version 2

Tohru Aomori, Koujirou Yamamoto, Atsuko Oguchi-Katayama, Yuki Kawai, Takefumi Ishidao, Yasumasa Mitani, Yasushi Kogo, Alexander Lezhava, Yukiyoshi Fujita, Kyoko Obayashi, Katsunori Nakamura, Hugo Kohnke, Mia Wadelius, Lena Ekströ, Cristine Skogastierna, Anders Rane, Masahiko Kurabayashi, Masami Murakami, Paul E. Cizdziel, Yoshihide HayashizakiRyuya Horiuchi

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Background: Polymorphisms of the CYP2C9 (cytochrome P450, family 2, subfamily C, polypeptide 9) gene (CYP2C92, CYP2C93) and the VKORC1 (vitamin K epoxide reductase complex, subunit 1) gene (-1639G>A) greatly impact the maintenance dose for the drug warfarin. Prescreening patients for their genotypes before prescribing the drug facilitates a faster individualized determination of the proper maintenance dose, minimizing the risk for adverse reaction and reoccurrence of thromboembolic episodes. With current methodologies, therapy can be delayed by several hours to 1 day if genotyping is to determine the loading dose. A simpler and more rapid genotyping method is required. methods: We developed a single-nucleotide polymorphism (SNP)-detection assay based on the SMart Amplification Process version 2 (SMAP 2) to analyze CYP2C92, CYP2C93, and VKORC1-1639G>A polymorphisms. Blood from consenting participants was used directly in a closed-tube real-time assay without DNA purification to obtain results within 1 h after blood collection. results: We analyzed 125 blood samples by both SMAP 2 and PCR-RFLP methods. The results showed perfect concordance. CONCLUSIONS: The results validate the accuracy of the SMAP 2 for determination of SNPs critical to personalized warfarin therapy. SMAP 2 offers speed, simplicity of sample preparation, the convenience of isothermal amplification, and assay-design flexibility, which are significant advantages over conventional genotyping technologies. In this example and other clinical scenarios in which genetic testing is required for immediate and better-informed therapeutic decisions, SMAP 2-based diagnostics have key advantages.

Original languageEnglish
Pages (from-to)804-812
Number of pages9
JournalClinical Chemistry
Volume55
Issue number4
DOIs
Publication statusPublished - 2009 Apr 1

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

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    Aomori, T., Yamamoto, K., Oguchi-Katayama, A., Kawai, Y., Ishidao, T., Mitani, Y., Kogo, Y., Lezhava, A., Fujita, Y., Obayashi, K., Nakamura, K., Kohnke, H., Wadelius, M., Ekströ, L., Skogastierna, C., Rane, A., Kurabayashi, M., Murakami, M., Cizdziel, P. E., ... Horiuchi, R. (2009). Rapid single-nucleotide Polymorphism detection of cytochrome P450 (CYP2C9) and vitamin k epoxide reductase (VKORC1) genes for the warfarin dose adjustment by the Smart-Amplification Process Version 2. Clinical Chemistry, 55(4), 804-812. https://doi.org/10.1373/clinchem.2008.115295