Detection of human papillomaviruses

• US 6,902,899 B2
• Filed: 08/22/2002
• Issued: 06/07/2005
• Est. Priority Date: 02/26/2000
• Status: Expired due to Fees

• Current Assignee: Eberhard-Karls-Universitat Tubingen Universitatsklinikum
• Original Assignee: Eberhard-Karls-Universitat Tubingen Universitatsklinikum
• Inventors: Iftner, Thomas
• Primary Examiner(s): SWITZER, JULIET CAROLINE
• Application Number:
  US10/227,638
• Publication Number:
  US 20030129585A1
• Time in Patent Office: 1,020 Days
• Field of Search: 435/6, 435/91.2, 536/24.3, 536/24.32
• US Class Current: 435/7.24
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6851 Quantitative amplification
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/708 for papilloma

Abstract

A method for detecting human papillomaviruses (HPV) in biological material is described, in which method nucleic acids are extracted/isolated from biological material and HPV-specific DNA is detected in the isolated nucleic acids. This method uses at least one of the nucleotide sequences SEQ ID No. 1 or SEQ ID No. 2 from the enclosed sequence listing or sequences which bind to sequences to which one of the sequences SEQ ID No. 1 or SEQ ID No. 2 binds or sequences which are complementary to the abovementioned sequences.

Claims

1. A method for detecting human papillomaviruses (HPV) in biological material, comprising the steps of:

(a) extracting and isolating nucleic acids from biological material, and

(b) detecting HPV-specific DNA in the isolated nucleic acids, by detecting specific HPV DNA segments using a pair of oligonucleotides selected from the group consisting of the pair SEQ ID NO;

1 and SEQ ID NO;

2, or the complement thereof or the pair SEQ ID NO;

1 and SEQ ID NO;

3 or the complement thereof.

2. The method of claim 1, wherein the detection is effected by way of amplifying the specific HPV-DNA segments by means of a polymerase chain reaction (PCR).

3. The method of claim 2, wherein the PCR is characterized by the following temperature profile:

Heating;

 

5-15 min., 90-100°

C.;

Heat denaturation;

30-90 sec., 90-100°

C.;

Hybridization;

30-90 sec., 50-60°

C.;

Polymerization;

30-180 sec., 67-77°

C.;

Cooling;

 

2-10 min., 67-77°

C.

4. The method of claim 2, wherein the primer pair employed for the PCR comprises a pair of oligonucleotides consisting of SEQ ID NO:

1 and SEQ ID NO;

2.

5. The method of claim 3, wherein the primer pair employed for the PCR comprises a pair of oligonucleotides consisting of SEQ ID NO:

1 and SEQ ID NO;

2.

6. The method of claim 2, wherein the primer pair employed for the PCR comprises a pair of oligonucleotides consisting of SEQ ID NO:

1 and SEQ ID NO;

3.

7. The method of claim 3, wherein the primer pair employed for the PCR comprises a pair of oligonucleotides consisting of SEQ ID NO:

1 and SEQ ID NO;

3.

8. The method of claim 1, wherein the HPVs are typed by sequencing the detected HPV DNA segments.

9. The method of claim 2, wherein the HPVs are typed by sequencing the amplified HPV DNA segments.

10. The method of claim 3, wherein the HPVs are typed by sequencing amplified HPV DNA segments.