title: CYP2A6 genotyping methods and strategies using real-time and end point PCR platforms.
abstract: CYP2A6 genotyping is of clinical importance--CYP2A6 gene variants influence nicotine metabolism and are associated with nicotine dependence, cigarettes per day, smoking cessation and the risk for tobacco-associated cancers. CYP2A6 gene variants also influence the metabolism of therapeutic drugs, such as the anticancer agents, tegafur and letrozole. Over the years, CYP2A6 genotyping methods have evolved to incorporate novel gene variants and to circumvent genotyping errors resulting from the high degree of homology between CYP2A6 and neighboring CYP2A genes. Herein, CYP2A6 genotyping strategies are described for commonly genotyped functionally significant alleles including SNPs, small insertions/deletions and more complex structural variants. The methods presented utilize higher throughput SYBR green real-time PCR technology in addition to standard thermocycling.
authors.affiliation: Department of Pharmacology & Toxicology, University of Toronto, 1 King's College Circle, Toronto, M5S 1A8 ON, Canada.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Wassenaar
authors.affiliation: Department of Pharmacology & Toxicology, University of Toronto, 1 King's College Circle, Toronto, M5S 1A8 ON, Canada.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Campbell Family Mental Health Research Institute, Centre for Addiction & Mental Health (CAMH), Departments of Pharmacology & Toxicology & Psychiatry, University of Toronto, 1 King's College Circle, Toronto, M5S 1A8 ON, Canada.
authors.forename: Rachel F
authors.initials: RF
authors.lastname: Tyndale
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics*; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D054643:INDEL Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D013250:Steroid Hydroxylases / Q000235:genetics*
chemical_list: D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; C029009:steroid hormone 7-alpha-hydroxylase
keywords: CYP2A6; SNP; end point PCR; gene deletion; hybrid allele; real-time PCR