title: Pyrosequencing-based screening for genetic polymorphisms in cytochrome P450 2B6 of potential clinical relevance.
abstract: OBJECTIVE
  Three exonic single nucleotide polymorphisms (SNPs) in the cytochrome P450 2B6 (CYP2B6) gene, 516G>T, 785A>G and 1459C>T, have been described to be associated with functional changes in the CYP2B6 catalytic activity or protein expression. They are therefore of potential clinical importance for drug efficacy and safety of CYP2B6 substrates, in particular of antiretroviral therapy regimes that include efavirenz. Therefore, we aimed at providing genetic screening assays for these three SNPs.
  
  
  METHODS
  Simplex Pyrosequencing assays were developed for the selected CYP2B6 SNPs. A total of 273 DNA samples from healthy volunteers of Caucasian ethnicity were genotyped.
  
  
  RESULTS
  The presently developed Pyrosequencing assays afford an accurate determination of the three SNPs in the 273 DNA samples. The assays were verified by routine implementation of control samples obtained by conventional sequencing. The frequencies for the variant alleles 516T, 785G and 1459T were 0.22, 0.24 and 0.13, respectively. All genotype frequencies were in agreement with the Hardy-Weinberg equilibrium. CYP2B6 alleles CYP2B6*5A (1459T), CYP2B6*6 (516T/785G) and *7B (516T/785G/1459T) were found at allelic frequencies of 0.12, 0.20 and 0.01.
  
  
  CONCLUSIONS
  The present reliable and quick Pyrosequencing assays afford facilitating future research on the importance of CYP2B6 pharmacogenetics for personalized drug therapy with CYP2B6 substrates.
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Institute of Clinical Pharmacology, Johann Wolfgang Goethe-University, Theodor Stern Kai 7, D-60590 Frankfurt, Germany.
authors.forename: Maren
authors.initials: M
authors.lastname: Rohrbacher
authors.forename: Anja
authors.initials: A
authors.lastname: Kirchhof
authors.forename: Gerd
authors.initials: G
authors.lastname: Geisslinger
authors.forename: Jörn
authors.initials: J
authors.lastname: Lötsch
mesh_terms: D000328:Adult; D000480:Alkynes; D000483:Alleles; D019380:Anti-HIV Agents / Q000493:pharmacokinetics* / Q000627:therapeutic use*; D023241:Antiretroviral Therapy, Highly Active; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics*; D048588:Benzoxazines; D003521:Cyclopropanes; D065702:Cytochrome P-450 CYP2B6; D005006:Ethnicity; D005091:Exons / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010078:Oxazines / Q000493:pharmacokinetics* / Q000627:therapeutic use*; D010089:Oxidoreductases, N-Demethylating / Q000235:genetics*; D011110:Polymorphism, Genetic / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000295:instrumentation*
chemical_list: D000480:Alkynes; D019380:Anti-HIV Agents; D048588:Benzoxazines; D003521:Cyclopropanes; D010078:Oxazines; D001189:Aryl Hydrocarbon Hydroxylases; C585599:CYP2B6 protein, human; D065702:Cytochrome P-450 CYP2B6; D010089:Oxidoreductases, N-Demethylating; C098320:efavirenz