Methods and compositions for detecting dihydropyrimidine dehydrogenase splicing mutations

Abstract

The present invention provides methods, compositions and kits for the detection of genetic polymorphisms or mutations of the dihydropyrimidine dehydrogenase deficiency (DPDD). The polymorphisms or mutations generally occur in the dihydropyrimidine dehydrogenase (DPD) gene in chromosome 1. Also provided are mutant forms of DPD.

1 - 14 . (canceled) 
     
     
         15 . A kit comprising a container, a first PCR primer which binds to DNA 3′ of a splice site in tie genomic DNA for dihydropyrimidine dehydrogenase gene for an exon encoding amino acids 581-635, and a second PCR primer which binds to DNA 5′ of a splice site in the genomic DNA for dihydropyrimidine dehydrogenase gene for an exon encoding amino acids 581-635. 
     
     
         16 . The kit of  claim 15  (original), wherein the kit further comprises instructions for the detection of splicing site defects in the dihydropyrimidine dehydrogenase gene. 
     
     
         17 . The kit of  claim 15 , wherein the kit further comprises Mae II. 
     
     
         18 . The kit of  claim 15 , wherein the first PCR primer hybridizes under stringent conditions to a nucleic acid complementary to DELF1. 
     
     
         19 . The kit of  claim 15 , wherein the second PCR primer hybridizes under stringent conditions to a nucleic acid complementary to DELR1. 
     
     
         20 . The kit of  claim 15 , further comprising a restriction endonuclease which recognizes a Mae II cleavage site. 
     
     
         21 . The kit of  claim 15 , wherein the first PCR primer binds to intronic dihydropyrimidine dehydrogenase DNA. 
     
     
         22 . The kit of  claim 15 , wherein the second PCR primer binds to intronic dihydropyrimidine dehydrogenase DNA. 
     
     
         23 . The kit of  claim 15 , wherein the intronic dihydropyrimidine dehydrogenase genomic DNA flanks the exon which encodes amino acids 581-635.