Methods and compositions for detecting dihydropyrimidine dehydrogenase splicing mutations
AssigneeThe United States of America as represented by the Department of Health and Human Services
InventorsFrank J. Gonzalez, Pedro Fernandez-Salguero
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Abstract
The present invention provides compositions, methods and kits for the detection of genetic polymorphisms or mutations related to dihydropyrimidine dehydrogenase deficiency (DPDD). The polymorphism or mutations generally occur in the dihydropyrimidine dehydrogenase DPD gene in chromosome 1. Also provided are mutant forms of DPD.
Claims18
A method of screening for a splicing defect in a human dihydropyrimidine dehydrogenase gene, wherein said gene comprises the sequence of SEQ ID NO:
- 1 or the variant thereof wherein the residue at position 434 is an A and wherein said defect causes the exon of SEQ ID NO:
- 1 to be skipped, said method comprising determining whether said gene has an A residue or a G residue at said position 434, wherein the presence of an A residue, but not a G residue, at said position 434 indicates the presence of the splicing defect causing the exon of SEQ ID NO:1 to be skipped.
A method of screening a human for sensitivity to 5-fluorouracil, comprising:
- (a) isolating genomic DNA from the human, said genomic DNA comprising the sequence of SEQ ID NO:1 or comprising the sequence of SEQ ID NO:1 wherein the residue at position 434 is an A;
- (b) amplifying under stringent conditions a fragment of the genomic DNA comprising position 434 of SEQ ID NO:1;
- and
- (c) determining whether the amplified fragment has an A or G residue at position 434;
- wherein the substitution of an A residue by a G residue at position 434 indicates the patient is sensitive to 5-fluorouracil.
A composition comprising a polymerase chain reaction primer from about 15 to about 20 nucleotides long wherein the nucleotide sequence of said primer is exactly complementary to a nucleotide sequence of SEQ ID NO:
- 1 between position 434 and position 861.
A method of testing a human for a single nucleotide polymorphism at position 434 of SEQ ID NO:1, the method comprising:
- a) isolating a sample from the human of genomic DNA wherein the genomic DNA comprises the nucleotide sequence of SEQ ID NO:1 or the variant thereof wherein the nucleotide corresponding to position 434 of SEQ ID NO:1 is A;
- b) selectively amplifying a fragment of the genomic DNA wherein the amplified fragment comprises position 434 of SEQ ID NO:1;
- and
- c) determining whether the residue at position 434 of SEQ ID NO:1 is an A or a G.
A composition comprising a polymerase chain reaction primer from about 15 to about 20 nucleotides long, wherein the nucleotide sequence of said primer is exactly complementary to a nucleotide sequence of SEQ ID NO:
- 1 between position 1 and position 268.
Dependencies are inferred from the claim text.