Genotyping assay to predict cytochrome P4503A5 exression

Abstract

Genetic polymorphisms responsible or associated with altered expression of cytochrome P450 CYP3A5 enzyme are described. Single nucleotide polymorphisms are provided. Methods for identifying subjects having a low or high drug metabolizing phenotype associated with CYP3A5 expression are provided. Assays, kits and methods for determining and assaying the CYP3A5 genotype and phenotype of individual patients are disclosed. Oligonucleotide probes and primers for use in the assays, kits and methods are described. Assays and methods for determining and evaluating an individual's metabolism of drugs and therapeutic agents, the potential for drug interactions, and thereby toxicity and effectiveness of certain drugs and treatment modalities, are provided.

1 - 32 . (canceled)  
     
     
         33 . A test kit suitable for determining cytochrome P450 3A5 (CYP3A5) genotype, and thereby determining expression of cytochrome P450 3A5 (CYP3A5) protein in an individual comprising: 
 (a) a predetermined amount of a first amplification primer complementary to a region 5′ to nucleotide 23 of SEQ ID NO:73 within intron 3 of the CYP3A5 gene;    (b) a predetermined amount of a second amplification primer complementary to a region 3′ to nucleotide 23 of SEQ ID NO:73 within intron 3 of the CYP3A5 gene:    (c) other reagents; and    (d) directions for use of said kit.    
     
     
         34 . The test kit of  claim 33  wherein the first amplification primer has the sequence corresponding to SEQ ID NO: 24 or SEQ ID NO: 26 and the second amplification primer has the sequence corresponding to SEQ ID NO: 25 or SEQ ID NO:27.  
     
     
         35 . A test kit suitable for determining cytochrome P450 3A5 (CYP3A5) genotype, and thereby determining expression of cytochrome P450 3A5 (CYP3A5) protein in an individual comprising: 
 (a) a predetermined amount of a first amplification primer complementary to a region 5′ to nucleotide 29 of SEQ ID NO:74 within exon 7 of the CYP3A5 gene;    (b) a predetermined amount of a second amplification primer complementary to a region 3′ to nucleotide 29 of SEQ ID NO:74 within exon 7 of the CYP3A5 gene:    (c) other reagents: and    (d) directions for use of said kit.    
     
     
         36 . The test kit of  claim 35  wherein the first amplification primer has the sequence corresponding to SEQ ID NO: 30 or SEQ ID NO: 31 and the second amplification primer has the sequence corresponding to SEQ ID NO: 16 or SEQ ID NO:32.  
     
     
         37 . An isolated oligonucleotide primer having a sequence selected from the group of SEQ ID NOS: 24-27 and 33 or a fragment thereof which is at least ten bases long.  
     
     
         38 . An isolated oligonucleotide primer having a sequence selected from the group of SEQ ID NOS: 16, 30-32 and 34 or a fragment thereof which is at least ten bases long.