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SIMULTANEOUS GENOTYPING OF CYP2D6*3 , *4 , *5 AND *6 POLYMORPHISMS IN A SPANISH POPULATION THROUGH MULTIPLEX LONG POLYMERASE CHAIN REACTION AND MINISEQUENCING MULTIPLEX SINGLE BASE EXTENSION ANALYSIS
Author(s) -
Crescenti A,
Mas S,
Gassó P,
Baiget M,
Bernardo M,
Lafuente A
Publication year - 2007
Publication title -
clinical and experimental pharmacology and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.752
H-Index - 103
eISSN - 1440-1681
pISSN - 0305-1870
DOI - 10.1111/j.1440-1681.2007.04665.x
Subject(s) - genotyping , genotype , allele , biology , genetics , multiplex polymerase chain reaction , multiplex , polymerase chain reaction , restriction fragment length polymorphism , cyp2d6 , population , allele frequency , microbiology and biotechnology , gene , medicine , environmental health
SUMMARY1 The aim of the present study was to perform a descriptive study of the prevalence of the four major CYP2D6 poor metaboliser (PM) alleles ( *3 , *4 , *5 and *6 ) in a Spanish population ( n = 290) using a method based on a new combination of multiplex long polymerase chain reaction (PCR) and minisequencing through multiplex single base extension (SBE) analysis. 2 The method was validated using different strategies, such as allelic discrimination assay and PCR–restriction fragment length polymorphism (RFLP). 3 The allele frequencies were similar to those described for other Spanish populations, namely 0.9% (95% confidence interval (CI) 0.5–1.3), 16.4% (95% CI 14.9–18.0), 2.7% (95% CI 2.0–3.4) and 0.7% (95% CI 0.3–1.0) for the *3 , *4 , *5 and *6 alleles, respectively. The results were satisfactory and left little doubt as to the genotypes, which were confirmed either by allelic discrimination assay ( *4 and *6 ) or PCR‐RFLP ( *3 ) with 100% concordance. 4 The present study corroborates the low prevalence of the most frequent polymorphism (CYP2D6 *4 ) that leads to null CYP2D6 activity in Spain and the allelic geographical gradient between Caucasian populations in the north and south. The present study reports a technique for the detection of four polymorphisms that account for 98% of the CYP2D6 defect alleles. This multiplex long PCR–SBE technique is a combination of several known methods to genotype CYP2D6 alleles ( *3 , *4 , *5 and *6 ). Given the importance of CYP2D6 in drug metabolism and the need to genotype a large number of samples, we believe that this method will find broad application.