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


METHODS
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.


RESULTS
We tested the multiplex in 152 samples and found it to be 100% concordant with the results of other methods.


CONCLUSIONS
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.
Laboratory of Forensic Genetics and Population Genetics, Department of Toxicology and Health Sanitary, Faculty of Medicine, Complutense University of Madrid, 28040 Madrid, Spain. amlopezparra@med.ucm.es
A M
AM
López-Parra
A M
AM
Borobia
C
C
Baeza
E
E
Arroyo-Pardo
A J
AJ
Carcas
D000074:Acenocoumarol / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D000925:Anticoagulants / Q000008:administration & dosage / Q000493:pharmacokinetics; D001057:Apolipoproteins E / Q000235:genetics*; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics*; D065729:Cytochrome P-450 CYP2C9; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D000072470:Cytochrome P450 Family 4; D006801:Humans; D006899:Mixed Function Oxygenases / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity; D014631:Valerates; D064417:Vitamin K Epoxide Reductases
D000925:Anticoagulants; D001057:Apolipoproteins E; D014631:Valerates; C012093:Validol; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9; D001189:Aryl Hydrocarbon Hydroxylases; D000072470:Cytochrome P450 Family 4; C117480:CYP4F2 protein, human; C578523:VKORC1 protein, human; D064417:Vitamin K Epoxide Reductases; D000074:Acenocoumarol