Method of determining Dihydropyrimidine dehydrogenase gene expression
AssigneeResponse Genetics, Inc.
InventorKathleen D. Danenberg
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Abstract
The present invention relates to prognostic methods which are useful in medicine, particularly cancer chemotherapy. The object of the invention to provide a method for assessing Dihydropyrimidine dehydrogenase (DPD) expression levels in tissues and prognosticate the probable resistance of a patient's tumor to treatment with 5-FU based therapies by examination of the amount of DPD mRNA in a patient's tumor cells and comparing it to a predetermined threshold expression level. More specifically, the invention provides to oligonucleotide primer pairs DPD3A and DPD3B and methods comprising their use for detecting levels of Dihydropyrimidine dehydrogenase (DPD) mRNA.
Claims26
An oligonucleotide having the sequence of SEQ ID NO:1 or which is substantially identical thereto.
An oligonucleotide having the sequence of SEQ ID NO:2 or which is substantially identical thereto.
An oligonucleotide having the sequence of SEQ ID NO:7 or which is substantially identically thereto.
An oligonucleotide having the sequence of SEQ ID NO:8 or which is substantially identically thereto.
A kit for detecting expression of a Dihydropyrimidine dehydrogenase (DPD) gene in a tissue obtained from a patient comprising oligonucleotide pair DPD3A or a pair of oligonucleotides substantially identical thereto or oligonucleotide pair DPD3B or a pair of oligonucleotides substantially identical thereto.
A method of determining the relative level of Dihydropyrimidine dehydrogenase (DPD) gene expression in a tissue sample comprising:
- (a) obtaining a tumor sample from a patient;
- (b) isolating mRNA from said tumor sample;
- (c) amplifying the mRNA using an oligonucleotide primer having the sequence of SEQ ID:
- 1, or which is substantially identical thereto and an oligonucleotide having the sequence SEQ ID:
- 2, or which is substantially identical thereto;
- (d) comparing the amount of Dihydropyrimidine dehydrogenase (DPD) mRNA from step (c) to an amount of mRNA of an internal control gene.
A method for determining a 5-Fluorouracil-based chemotherapeutic regimen for treating a tumor in patient comprising:
- (a) obtaining a tumor sample from the patient;
- (b) isolating mRNA from said tumor sample;
- (c) subjecting the mRNA to amplification using a pair of oligonucleotide primers having the sequence of SEQ ID:
- 1, or which is substantially identical thereto and an oligonucleotide having the sequence SEQ ID:
- 2, or which is substantially identical thereto to obtain an amplified sample,
- (d) determining the amount of Dihydropyrimidine dehydrogenase (DPD) mRNA in the amplified sample;
- (e) comparing the amount of Dihydropyrimidine dehydrogenase (DPD) mRNA in the amplified sample with a predetermined threshold level for DPD expression;
- (f) determining a 5-Fluorouracil-based chemotherapeutic regimen for said patient based on the difference in amount of DPD mRNA in the amplified sample and the threshold level for DPD gene expression.
A method of determining the relative level of Dihydropyrimidine dehydrogenase (DPD) gene expression in a tissue sample comprising;
- (a) obtaining a tumor sample from a patient;
- (b) isolating mRNA from said tumor sample;
- (c) amplifying the mRNA using an oligonucleotide primer having the sequence of SEQ ID:
- 7, or which is substantially identical thereto and an oligonucleotide having the sequence SEQ ID:
- 8, or which is substantially identical thereto;
- (d) comparing the amount of the mRNA from step (c) to an amount of mRNA of an internal control.
A method for determining a 5-Fluorouracil-based chemotherapeutic regimen for treating a tumor in a patient comprising:
- (a) obtaining a tumor sample from the tumor;
- (b) isolating mRNA from a tumor sample;
- (c) subjecting the mRNA to amplification using a pair of oligonucleotide primers having of the sequence of SEQ ID:
- 7, or which is substantially identical thereto and an oligonucleotide having the sequence SEQ ID:
- 8, or which is substantially identical thereto, to obtain an amplified sample;
- (d) determining the amount of Dihydropyrimidine dehydrogenase (DPD) mRNA in the amplified sample;
- (e) comparing the amount of Dihydropyrimidine dehydrogenase (DPD) mRNA in the amplified sample with a predetermined threshold level for DPD expression;
- (f) determining a 5-Fluorouracil-based chemotherapeutic regimen for said patient based on the difference in amount of DPD mRNA in the amplified sample and the threshold level for DPD gene expression.
Dependencies are inferred from the claim text.