IDENTIFICATION OF HIGH GRADE OR >= CIN2 FOR EARLY STAGES AND LATE STAGES DETECTION, SCREENING, AND DIAGNOSIS OF HUMAN PAPILLOMAVIRUS (HPV) AND HPV-ASSOCIATED CANCERS

• US 20120052486A1
• Filed: 05/06/2010
• Published: 03/01/2012
• Est. Priority Date: 11/13/2006
• Status: Active Grant

• Current Assignee: Heer Medical Technology Development Co., Ltd.
• Original Assignee: Onco Health Corporation
• Inventors: Cheng, Shuling
• Granted Patent: US 8,916,342 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: G01N 2333/025 Papovaviridae, e.g. papillomavirus, polyomavirus, SV40, BK virus, JC virus
• CPC Class Codes: G01N 2800/56 Staging of a disease; Further complications associated with the disease
• CPC Class Codes: G01N 33/56983 Viruses
• CPC Class Codes: G01N 33/57411 of cervix

Abstract

A method and a kit for determining a disease stage of a papilloma virus infection in a human subject is disclosed. The method comprises: obtaining a sample from said human subject, said sample consisting of a tissue sample comprising proteins or a cell sample comprising proteins; contacting said sample with one or more antibodies that specifically bind to one or more recombinant HPV proteins; determining a presence, absence, or amount of at least one protein in said sample that specifically binds to said antibodies and determining said disease stage of papilloma virus infection in said human subject based on said determined presence, absence, or amount of said one or more proteins.

Claims

1. A method for determining a disease stage of papilloma virus infection in a human subject comprising:

receiving a sample obtained from said human subject, said sample consisting of a tissue sample comprising proteins or a cell sample comprising proteins;

contacting said sample with a first antibody that specifically binds to a recombinant HPV protein, where in

the recombinant HPV protein is selected from the group consisting of a recombinant papillomavirus E6 gene product, a recombinant papillomavirus E7 gene product, a recombinant papillomavirus L1 gene product, a recombinant papillomavirus truncated L1 gene product, and a recombinant papillomavirus L2 gene product, wherein said contacting takes place under conditions that promote specific binding of said antibody;

determining presence, absence, or an amount of a first protein in said sample that specifically binds to the first antibody based at least in part on a measure of specific binding of the first antibody to the sample contacted by the antibody; and

determining said disease stage of papilloma virus infection in said human subject based on said determined presence, absence, or amount of said one or more proteins in said sample.

2. The method of claim 1 wherein said first one or more proteins comprise one or more proteins selected from the group consisting of a papillomavirus E6 gene product, a papillomavirus E7 gene product, a papillomavirus L1 gene product, a papillomavirus truncated L1 gene product, and a papillomavirus L2 gene product.

3. The method of claim 2 wherein said first one or more proteins comprise one or more proteins selected from the group consisting of a papillomavirus E6 gene product and a papillomavirus E7 gene product.

4. The method of claim 1, further comprising:

contacting said sample with a second antibody;

determining presence, absence or an amount of a second one or more proteins selected from the group consisting of p16INK4a, pRB, p53, E2F, E2F activated cell cycle protein, cyclin dependent kinase, CDK4, CDK6, Ki-67 (MIB-1), MYC protein, cyclin-A, cyclin-B, cyclin-E, telomerase-TERC, MCM2, TOP2A, heat shock protein 40 (HSP₄₀), heat shock protein 60 (HSP₆₀), heat shock protein 70 (HSP₇₀), CA9/MN, laminin5, brn-3a, CDK N2, topoisomerase 2A, microsome maintenance protein-2, microsome maintenance protein-4, microsome maintenance protein-5, survivin, VEGF, p27 (kip1), and p21 (waf) in said sample based on a measure of specific binding of the second antibody to said contacted sample; and

wherein said determining the disease state for the human subject for papillomavirus infection is further based on said determined presence, absence, or amount of said second protein.

5. The method of claim 4 wherein said one or more proteins comprises p16^INK4a.

6. The method of claim 1 wherein said sample is in situ.

7. The method of claim 1 wherein said sample comprises a tissue sample, and said determining comprises a immunohistochemistry assay.

8. The method of claim 1 wherein said sample comprises a cell sample, and said determining comprises an immunocytochemistry assay.

9. The method of claim 1 wherein said determining said disease stage comprises determining the presence or absence of HSIL.

10. The method of claim 1 wherein said determining said disease stage comprises determining the presence or absence of late stage HPV infection.

11. The method of claim 1 wherein said determining said disease stage comprises determining the presence or absence of high grade dysplasia.

12. The method of claim 1 wherein said determining said disease stage comprises determining the presence or absence of a disease stage≧

CIN2.

13. The method of claim 1 where in said disease stage is ≧

CIN2 or <

CIN2.

14. The method of claim 13 wherein said ≧

CIN2 is CIN2/3.

15. The method of claim 1 wherein said disease stage is HSIL.

16. The method of claim 1 wherein said disease stage is late stage HPV infection.

17. The method of claim 1 wherein said disease stage is high grade dysplasia.

19. The method of claim 1 wherein said papillomavirus comprises one or more viruses selected from the group consisting of HPV-6, HPV-11, HPV-16, HPV-18, HPV-31, HPV-33, HPV-35, HPV-39, HPV-42, HPV-43, HPV-44, HPV-45, HPV-51, HPV-52, HPV-53, HPV-54, HPV-55, HPV-56, HPV-58, HPV-59, and HPV-66.

20. The method of claim 19 wherein said papillomavirus comprises one or more viruses selected from the group consisting of HPV-16 and HPV-18.

21. The method of claim 1 wherein said recombinant papillomavirus E6 gene product is a recombinant HPV-16 E6 gene product or a recombinant HPV-18 E6 gene product.

22. The method of claim 1 wherein said recombinant papillomavirus E7 gene product is a recombinant HPV-16 E7 gene product or a recombinant HPV-18 E7 gene product.

23. The method of claim 1 wherein said cell sample comprises normal cells, ASC-US cells, ASC-H cells, LSIL cells, HSIL cells, ADC cells, AGC cells or SCC cells.

24. The method of claim 1 wherein said tissue sample is benign, CIN1, CIN2, CIN3, SCC, AGC or ADC.

25. The method of claim 1 further comprising:

obtaining a second sample from said human subject, said sample comprising a cell sample; and

performing a cytological Papanicolaou smear assay on said sample.

26. The method of claim 1 further comprising:

obtaining a second sample from said human subject, said sample comprising a nucleic acid; and

performing a nucleic acid hybridization assay for human papillomavirus on said sample.

27. A kit comprising:

reagents to perform the method of claim 1; and

instructions for performing the method.

18. (canceled)