Methods and compositions for the detection of cervical disease

• US 7,157,233 B2
• Filed: 03/23/2005
• Issued: 01/02/2007
• Est. Priority Date: 03/24/2004
• Status: Active Grant

• Current Assignee: Tripath Imaging Incorporated (Becton, Dickinson & Co)
• Original Assignee: Tripath Imaging Incorporated (Becton, Dickinson & Co)
• Inventors: Fischer, Timothy J., Malinowski, Douglas P., Taylor, Adriann J., Parker, Margaret R.
• Primary Examiner(s): SALIMI, ALI REZA
• Application Number:
  US11/087,227
• Publication Number:
  US 20050260566A1
• Time in Patent Office: 650 Days
• Field of Search: 435/6, 435/7.1, 435/5, 435/345
• US Class Current: 435/6.12
• CPC Class Codes: C07K 16/3069 Reproductive system, e.g. ovaria, uterus, testes, prostate
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/112 Disease subtyping, staging or classification
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 2333/025 Papovaviridae, e.g. papillomavirus, polyomavirus, SV40, BK virus, JC virus
• CPC Class Codes: G01N 33/57411 of cervix
• CPC Class Codes: G01N 33/6875 Nucleoproteins

Abstract

Methods and compositions for identifying high-grade cervical disease in a patient sample are provided. The methods of the invention comprise detecting overexpression of at least one biomarker in a body sample, wherein the biomarker is selectively overexpressed in high-grade cervical disease. In particular claims, the body sample is a cervical smear or monolayer of cervical cells. The biomarkers of the invention include genes and proteins that are involved in cell cycle regulation, signal transduction, and DNA replication and transcription. In particular claims, the biomarker is an S-phase gene. In some aspects of the invention, overexpression of a biomarker of interest is detected at the protein level using biomarker-specific antibodies or at the nucleic acid level using nucleic acid hybridization techniques. Kits for practicing the methods of the invention are further provided.

Claims

1. A method for diagnosing high-grade cervical disease in a patient independent of the patient'"'"'s HPV infection status, said method comprising:

a) obtaining a body sample from said patient;

b) contacting said sample with at least one antibody, wherein said antibody specifically binds to a nuclear biomarker protein that is selectively overexpressed in high-grade cervical disease, wherein said biomarker protein is selected from the group consisting of p14ARF, p21^waf1, Topo2A, and Cyclin E; and

,

c) detecting binding of said antibody to said nuclear biomarker protein to determine if said biomarker protein is overexpressed in said sample, and thereby diagnosing high-grade cervical disease in the patient.

2. The method of claim 1, wherein the method comprises performing immunocytochemistry.

3. The method of claim 2, wherein the method is performed manually.

4. The method of claim 2, wherein the method is performed in an automated manner.

5. The method of claim 1, wherein the sample comprises cervical cells.

6. The method of claim 5, wherein the sample comprises a monolayer of cervical cells.

7. The method of claim 1, wherein the sensitivity of said method for diagnosing high-grade cervical disease is at least 70%.

8. The method of claim 1, wherein the specificity of said method for diagnosing high-grade cervical disease is at least 85%.

9. The method of claim 1, wherein the method is performed in response to the patient having an abnormal Pap smear result.

10. The method of claim 1, wherein the method is performed as a primary screen for high-grade cervical disease in a general patient population.

11. The method of claim 1 further comprising Papanicolaou (Pap) staining of the sample.