Methods and compositions for the detection of cervical disease

• US 7,455,973 B2
• Filed: 08/17/2006
• Issued: 11/25/2008
• 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:
  US11/506,012
• Publication Number:
  US 20060286595A1
• Time in Patent Office: 831 Days
• Field of Search: 435/6, 435/69.1
• 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 comprising detecting overexpression of at least one nucleic acid molecule that encodes a nuclear biomarker that is selectively overexpressed in high-grade cervical disease, wherein the nuclear biomarker is selected from the group consisting of p14ARF, p21^waf1, Topo2A, and Cyclin E, the method comprising:

(a) obtaining a body sample from the patient;

(b) isolating nucleic acid material from the sample;

(c) mixing said nucleic acid material with at least one pair of oligonucleotide primers specific for the nuclear biomarker and a thermostable DNA polymerase under conditions that are suitable for amplification by polymerase chain reaction (PCR);

d) performing PCR; and

,

e) detecting PCR amplification products.

2. The method of claim 1, wherein performing PCR comprises performing RT-PCR.

3. The method of claim 1, wherein the method is performed in conjunction with morphological analysis.

4. The method of claim 1, wherein the sample comprises cervical cells in suspension in a liquid-based preparation.

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

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

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

8. The method of claim 1, wherein the method if performed in conjunction with Papanicolaou (Pap) staining of a cervical sample from the patient.