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Last Updated: March 29, 2024

Claims for Patent: 6,277,581


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Summary for Patent: 6,277,581
Title: ODC allelic analysis method for assessing carcinogenic susceptibility
Abstract:The invention includes kits and methods for assessing the susceptibility of a mammal such as a human for carcinogenesis. The methods comprise determining whether the mammal comprises a certain allele of the mammal\'s odc gene. The methods include use of a probe which binds specifically with a portion of one allele of the odc gene and which comprises a fluorescent label and a fluorescence quencher. The methods also include use of such a probe and a polymerase enzyme for amplifying a portion of the odc gene, the polymerase having exonuclease activity whereby the probe can be nucleolytically degraded.
Inventor(s): O\'Brien; Thomas G. (Drexel Hill, PA), Guo; Yong Jun (Drexel Hill, PA)
Assignee: Lankenau Medical Research Center (Wynnewood, PA)
Application Number:09/516,357
Patent Claims:1. A method of assessing the relative susceptibility of a mammal to an epithelial cancer, the method comprising determining whether the mammal comprises an A-allele of the odc gene, whereby if the mammal comprises the A-allele, then the mammal has a greater susceptibility to the epithelial cancer than a mammal of the same type which does not comprise the A-allele.

2. The method of claim 1, wherein the mammal is a human.

3. The method of claim 1, wherein the epithelial cancer is selected from the group consisting of a skin cancer, a cancer of the digestive system, an esophageal cancer, a gastric cancers, a colon cancer, a prostate cancer, a breast cancer, an hematopoietic cancer, a lung cancer, a melanoma, and a cervical cancer.

4. The method of claim 3, wherein the skin cancer is squamous cell carcinoma.

5. The method of claim 1, wherein determining whether the mammal comprises an A-allele of the odc gene comprises amplifying a reference portion of the mammal's genome.

6. The method of claim 5, wherein the reference portion is amplified using a pair of primers having nucleotide sequences SEQ ID NO: 3 and SEQ ID NO: 4.

7. The method of claim 6, wherein the reference portion is further amplified using a pair of nested primers having nucleotide sequences SEQ ID NO: 5 and SEQ ID NO: 6.

8. The method of claim 5, wherein the reference portion comprises a region of the odc gene.

9. The method of claim 5, wherein the reference portion comprises nucleotide residues +282 to +340 relative to the transcription start site.

10. The method of claim 5, wherein the reference portion has a nucleotide sequence selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

11. The method of claim 8, wherein the region comprises intron 1 of the odc gene.

12. The method of claim 8, wherein the region comprises the nucleotide residue located at position +321 relative to the transcription start site.

13. The method of claim 5, further comprising annealing an oligonucleotide probe with a target portion of the mammal's genome prior to amplifying the reference portion, wherein the target portion includes the nucleotide residue located at position +321 relative to the transcription start site of the odc gene.

14. The method of claim 13, wherein the probe comprises a fluorescent label.

15. The method of claim 14, wherein the fluorescent label is selected from the group consisting of FAM, TET, rhodamine, VIC, JOE, and HEX.

16. The method of claim 14, wherein the probe further comprises a fluorescence quencher.

17. The method of claim 16, wherein the quencher is selected from the group consisting of TAMRA and DABCYL.

18. The method of claim 17, wherein the probe has the nucleotide sequence SEQ ID NO: 14.

19. The method of claim 16, wherein one of the label and the quencher is attached to the probe within 10 nucleotide residues of the 3'-end of the probe and the other of the label and the quencher is attached to the probe within 10 nucleotide residues of the 5'-end of the probe.

20. The method of claim 16, wherein the reference portion is amplified using a DNA polymerase having 5'.fwdarw.3' exonuclease activity.

21. The method of claim 20, wherein the DNA polymerase is Thermus aquaticus DNA polymerase.

22. The method of claim 20, wherein the probe has a length from 15 to about 30 nucleotide residues.

23. The method of claim 22, wherein the size of the reference portion is not more than about 100 nucleotide residues.

24. The method of claim 1, wherein determining whether the mammal comprises an A-allele of the odc gene comprises contacting a polynucleotide derived from the mammal's genome with a molecular beacon probe, the probe having a targeting portion which is complementary to a target region of the odc gene, wherein the target region includes the nucleotide residue located at position +321 relative to the transcription start site of the odc gene.

25. The method of claim 24, wherein the targeting portion is completely complementary to the target region of the A-allele of the odc gene and has a length from about 20 to about 40 nucleotide residues.

26. The method of claim 25, further comprising contacting the polynucleotide with a second molecular beacon probe, the second probe having a targeting portion which is completely complementary to the target region of the G-allele and having a length from about 20 to about 40 nucleotide residues.

27. The method of claim 26, wherein the targeting portion of the first probe has the nucleotide sequence SEQ ID NO: 14 and the targeting portion of the second probe has the nucleotide sequence SEQ ID NO: 15.

28. The method of claim 24, wherein the target region comprises about 20 to about 30 consecutive nucleotide residues of a polynucleotide having a sequence selected from the group consisting of SEQ ID NOs: 1 and 2.

29. The method of claim 25, wherein the polynucleotide is selected from the group consisting of a chromosome of the mammal, a chromosomal fragment of the mammal, and an amplified portion of the genome of the mammal.

Details for Patent 6,277,581

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 06/04/1986 ⤷  Try a Trial 2019-03-01
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 ⤷  Try a Trial 2019-03-01
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b Injection 103132 ⤷  Try a Trial 2019-03-01
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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