Real-time polymerase chain reaction-based genotyping assay for single nucleotide polymorphism
AssigneeUniversity of Tennessee Research Foundation
InventorsCharles R. Yates, Duane Miller, Dick Gourley, Pengfei Song
Life of this patent
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Abstract
The present invention provides fluorescence-based real-time PCR assays for the rapid detection of single nucleotide polymorphisms (SNPs). The genotyping assay can be used to detect SNPs of a number of genes of interest that include, but are not limited to, the human multidrug resistance gene (MDR1) single nucleotide polymorphisms C3435T and G2677T, and cytochrome P-450 3A5 single nucleotide polymorphisms CYP3A5*3 (A22893G) and CYP3A5*6 (G30597A).
Claims4
A kit for genotyping human multidrug resistance gene (MDR1) single nucleotide polymorphism C3435T, comprising:
- a primer pair consisting of the primer sequences SEQ ID NO:
- 1 and SEQ ID NO:
- 3;
- and
- a primer pair consisting of the primer sequences SEQ ID NO:
- 2 and SEQ ID NO:
- 3.
A primer pair useful for genotyping human multidrug resistance gene (MDR1) single nucleotide polymorphism C3435T consisting of the primer sequences SEQ ID NO:
- 1 and SEQ ID NO:
- 3 or consisting of the primer sequences SEQ ID NO:
- 2 and SEQ ID NO:
- 3.
A method of genotyping human multidrug resistance gene (MDR1) single nucleotide polymorphism C3435T, comprising the steps of:
- preparing DNA samples from an individual;
- amplifying said DNA with a first primer pair comprising a wild type-specific primer and common reverse primer, wherein the first primer pair is a primer pair consisting of the primer sequences SEQ ID NO:
- 1 and SEQ ID NO:
- 3 and also amplifying said DNA with a second primer pair comprising a mutant-specific primer and common reverse primer, wherein the second primer pair is a primer pair consisting of the primer sequences SEQ ID NO:
- 2 and SEQ ID NO:
- 3 and
- identifying the products of said DNA amplification,
- wherein the presence of products amplified by said mutant-specific primer and said ieverse primer indicate said individual has at least one “T” allele at position 3435 of the MDR1 gene.
Dependencies are inferred from the claim text.