A multiplex assay to detect variations in the CYP2C9, VKORC1, CYP4F2 and APOE genes involved in acenocoumarol metabolism

• Published in: Clinical biochemistry Vol. 46; no. 1-2; pp. 167 - 169
• Main Authors: López-Parra, A.M., Borobia, A.M., Baeza, C., Arroyo-Pardo, E., Carcas, A.J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.01.2013
• Subjects: Acenocoumarol; Acenocoumarol - administration & dosage; Acenocoumarol - pharmacokinetics; Anticoagulants - administration & dosage; Anticoagulants - pharmacokinetics; APOE; Apolipoproteins E - genetics; Aryl Hydrocarbon Hydroxylases - genetics; CYP2C9; CYP4F2; Cytochrome P-450 CYP2C9; Cytochrome P-450 Enzyme System - genetics; Cytochrome P450 Family 4; Humans; Mixed Function Oxygenases - genetics; Multiplex Polymerase Chain Reaction - economics; Multiplex Polymerase Chain Reaction - methods; Polymorphism, Single Nucleotide; Sensitivity and Specificity; Valerates; Vitamin K Epoxide Reductases; VKORC1; VKORC1; CYP2C9; APOE; CYP4F2; Acenocoumarol
• ISSN: 0009-9120; 1873-2933
• DOI: 10.1016/j.clinbiochem.2012.08.005

We have developed a genotyping system to determine the alleles of genes related to interindividual variability in acenocoumarol dosage requirements. This genotyping system is intended for routine clinical use and therefore it is essential that it be simple, fast and inexpensive. We developed a PCR multiplex SNaPshot reaction that targets 6 SNPs (single nucleotide polymorphisms) in CYP2C9, CYP4F2, VKORC1 and APOE genes, which are associated with acenocoumarol dose requirements. We tested the multiplex in 152 samples and found it to be 100% concordant with the results of other methods. We successfully produced a reliable multiplex system for simultaneously typing 6 SNPs. This system may be used as a model for accurate, simple and inexpensive genotyping of SNPs related to dose requirements. This information allows the prediction of drug efficiency in patients prior to treatment with acenocoumarol and the prevention of adverse drug reactions. [Display omitted] ► We have developed a PCR multiplex SNaPshot reaction for CYP2C9, CYP4F2, VKORC1 and APOE. ► Polymorphism systems are related with appropriate dosage for acenocoumarol treatment. ► This SNaPshot assay is sensitive, simple, rapid and low cost. ► It can be included in the clinical routine prior to treatment with anticoagulants. ► This assay has been validated with other methodologies. (KASPar and TaqMan).