ワルファリン治療個別化に利用可能な簡易・同時遺伝子診断法の確立

Abstract

Identification of the gene encoding human vitamin K epoxide reductase complex subunit 1 (VKORC1 ),the main target molecule of coumarin derivatives,has contributed to clarifying racial differences in susceptibility to warfarin (Wf).However,although the discovery of genetic polymorphisms of the VKORC1 gene greatly aided understanding of the causes of great inter-individual variation in the response to Wf,a means of determining the optimal maintenance dose of the drug for each patient has not yet been established.From the viewpoint of application to both research and routine laboratory testing in the near future,we established a rapid and simple,simultaneous genotyping method for 2 known critical genetic factors,VKORC1 and CYP2C9 .The basic principle of this method is multi-primer PCR in which the genotypes of these genes are determined within 2 hr using a single thermal cycling condition.The intensity of the band amplified from the CYP2C9*3 allele is much greater than that obtained with the method currently used,indicating that the new method would be advantageous in visual diagnosis.Furthermore,since this method was applicable to non-blood derived samples,it would be suitable for a wide variety of patients whose blood is hard to collect,such as small children suffering from Kawasaki disease,or those with few leucocytes due to the adverse effects of chemotherapy with anti-cancer agents.