Method of identifying a polymorphism in CYP2D6
OwnerMarco Guida, Linda Benson +1inventors
InventorsMarco Guida, Linda Benson, Penelope Hopkins
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Abstract
The present invention relates to methods of detecting a polymorphism in a gene encoding xenobiotic metabolizing enzyme CYP2D6. The methodology disclosed is robust and can detect the presence of the wildtype and polymorphic sequences in the presence of gene duplication and gene conversion events.
Claims25
A method of determining a cytochrome P-450 2D6 genotype of an individual comprising:
- a) obtaining genomic DNA of the individual, wherein the genomic DNA comprises a CYP2D6 gene;
- b) subjecting a first portion of the genomic DNA to amplification conditions in the presence of a pair of primers, wherein one of the primers hybridizes to genomic DNA comprising a CYP2D6 exon 1 C188T polymorphism and does not hybridize to a CYP2D6 wild-type sequence at position 188 of exon 1 and wherein production of an amplification product indicates a CYP2D6*10 genotype.
An allele-specific amplification primer, wherein said primer hybridizes to, and primes amplification of, a fragment of a cytochrome P-450 2D6 gene comprising a CYP2D6 exon 1 C188T polymorphism but does not prime amplification of a cytochrome P-450 2D6 gene comprising the wildtype sequence at position 188.
A nucleic acid molecule, wherein said nucleic acid molecule is a fragment of a cytochrome P-450 2D6 gene comprising a CYP2D6 exon 1 C188T polymorphism wherein said nucleic acid molecule is between 10 and 50 nucleotides in length.
An amplification product containing the fragment of the CYP2D6 gene between nucleotide 68 and nucleotide 1212.
The amplification product produced by the method comprising:
- a) obtaining genomic DNA of an individual, wherein the genomic DNA comprises a CYP2D6 gene;
- b) subjecting at least a portion of the genomic DNA to amplification conditions in the presence of a pair of primers, wherein one of the primers hybridizes to genomic DNA comprising a CYP2D6 exon 1 C188T polymorphism and does not hybridize to a CYP2D6 wild-type sequence at position 188 of exon 1 to produce an amplification product.
The amplification product produced by the method comprising:
- a) obtaining genomic DNA of an individual, wherein the genomic DNA comprises a CYP2D6 gene;
- b) subjecting at least a portion of the genomic DNA to amplification conditions in the presence of a pair of primers, wherein one of the primers hybridizes to genomic DNA comprising a CYP2D6 wild-type sequence at position 188 of exon 1 and does not hybridize to a CYP2D6 exon 1 C188T polymorphism to produce an amplification product.
A method of prescribing a pharmaceutical composition to an individual comprising:
- a) obtaining genomic DNA of the individual, wherein the genomic DNA comprises a CYP2D6 gene;
- b) subjecting a first portion of the genomic DNA to amplification conditions in the presence of a pair of primers, wherein one of the primers hybridizes to genomic DNA comprising a CYP2D6 exon 1 C188T polymorphism and does not hybridize to a CYP2D6 wild-type sequence at position 188 of exon 1 and wherein production of an amplification product indicates a CYP2D6*10 genotype;
- c) subjecting a second portion of the genomic DNA to amplification conditions in the presence of a pair of primers, wherein one of the primers hybridizes to genomic DNA comprising a CYP2D6 wild-type sequence at position 188 of exon 1 and does not hybridize to a CYP2D6 exon 1 C188T polymorphism and wherein production of an amplification product indicates a CYP2D6 wild-type genotype;
- and,
- d) prescribing a pharmaceutical composition for the individual based on amplification results of steps (b) and (c).
Dependencies are inferred from the claim text.