Polymerase chain reaction/restriction fragment polymorphism method for the detection and typing of human papillomaviruses

• US 5,814,448 A
• Filed: 01/31/1996
• Issued: 09/29/1998
• Est. Priority Date: 07/19/1991
• Status: Expired due to Fees

• Current Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Original Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Inventors: Wright, Thomas C. Jr., Silverstein, Saul J., Lungu, Octavian
• Primary Examiner(s): Ketter, James
• Assistant Examiner(s): Brusca, John S.
• Application Number:
  US08/594,600
• Time in Patent Office: 972 Days
• Field of Search: 435/6, 435/91.2, 536/24.33, 536/23.72
• US Class Current: 435/6.12
• CPC Class Codes: C12Q 1/683 involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: C12Q 1/708 for papilloma
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

This invention provides a method of typing a human papillomavirus in a patient infected by human papillomavirus which comprises: obtaining a sample containing DNA from the human papillomavirus to be typed; amplifying the L1 portion of the human papillomavirus DNA; treating the resulting amplified DNA with a plurality of predetermined restriction enzymes so as to produce restriction fragments; and analyzing the fragments so produces so as to type the human papillomavirus.

Claims

1. A method of typing a human papillomavirus in a patient infected by human papillomavirus which comprises:

a. obtaining a sample containing DNA from the human papillomavirus to be typed;

b. amplifying the L1 portion of the human papillomavirus DNA;

c. treating the resulting amplified DNA with a plurality of predetermined restriction enzymes so as to produce restriction fragments; and

d. analyzing the fragments so produced so as to type the human papillomavirus.

2. The method of claim 1, wherein the patient has a cervical intraepithelial neoplasia.

3. The method of claim 1, wherein the patient has an invasive cervical carcinoma.

4. The method of claim 1, wherein the patient has a benign condylomata.

5. The method of claim 1, wherein the sample is infected cervical tissue.

6. The method of claim 1, wherein the sample is infected vaginal tissue.

7. The method of claim 1, wherein the sample is infected vulvar tissue.

8. The method of claim 1, wherein the sample is infected anogenital tissue.

9. The method of claim 1, wherein the amplification in step (b) is effected by means of the polymerase chain reaction using a plurality of sets of primers.

10. The method of claim 1, wherein the plurality of predetermined restriction enzymes in step (c) comprises Hae I, Pst I and Rsa I.

11. The method of claim 1, wherein the fragments are analyzed in step (d) by means of gel electrophoretic separation of the restriction fragments so produced.