Papillomavirus probe and a process for in vitro diagnosis of papillomavirus infections

• US 5,876,922 A
• Filed: 06/07/1995
• Issued: 03/02/1999
• Est. Priority Date: 07/31/1985
• Status: Expired due to Term

• Current Assignee: Roche Molecular Systems Inc (Roche Holding AG)
• Original Assignee: Institut National De La Sante Et De La Recherche Medicale (Inserm) (Allianz Global Investors GmbH), Institut Pasteur
• Inventors: Kremsdorf, Dina, Beaudenon, Sylvie, Croissant, Odile, Orth, Gerard, Pehau-Arnaudet, Gerard, Favre, Michel
• Primary Examiner(s): JONES, WILLIAM G
• Assistant Examiner(s): Tran, Paul B.
• Application Number:
  US08/483,165
• Time in Patent Office: 1,364 Days
• Field of Search: 435/6, 435/5, 435/252.3, 435/948, 536/23.72
• US Class Current: 435/5
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 2710/20022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12Q 1/708 for papilloma
• CPC Class Codes: Y10S 435/948 using viruses or cell lines

Abstract

The invention concerns human papillomavirus (HPV) DNA and more particularly the probes derived from these papillomaviruses, as well as the methods of detecting HPV using these probes. These human papillomaviruses are designated as HPV-2d, HPV-10b, HPV-14a, HPV-14b, HPV-15, HPV-17a, HPV-17b, HPV-19, HPV-20, HPV-21, HPV-22, HPV-23, HPV-24, HPV-28, HPV-29, HPV-31, HPV-32, HPV-IP2 and HPV-IP4.

Claims

1. A purified human papillomaviruses gene selected from the group consisting of E1, E6-E7, L1, and L2,

wherein said human papillomavirus is selected from the group consisting of HPV-2d, HPV-10b, HPV-14a, HPV-14b, HPV-15, HPV-17a, HPV-17b, HPV-19, HPV-20, HPV-21, HPV-22, HPV-23, HPV-24, HPV-28, HPV-29, HPV-31, HPV-32, HPV-IP2 and HPV-IP4.

3. Recombinant human papillomavirus (HPV) DNA comprising a gene according to claim 1.

4. A probe for the detection of human papillomavirus (HPV) DNA, said probe comprising a gene according to claim 1 or the purified DNA according to claim 2.

5. A method for the in vitro detection of infection due to a papillomavirus of the HPV IP4 type, said method comprising

placing the probe of claim 4 in contact with nucleic acids of a biological sample under stringent hybridization conditions;

detecting a hybrid formed between DNA in the sample and said probe;

and correlating the presence of said hybrid with an infection due to the presence of a papillomavirus of the HPV IP4 type.

2. A purified DNA, which hybridizes under stringent conditions to the DNA of human papillomavirus (HPV) selected from the group consisting of HPV-2d, HPV-10b, HPV-14a, HPV-14b, HPV-15, HPV-17a, HPV-17b, HPV-19, HPV-20, HPV-21, HPV-22, HPV-23, HPV-24, HPV-28, HPV-29, HPV-31, HPV-32, HPV-IP2 and HPV-IP4 wherein said stringent conditions comprise contacting said purified DNA with said HPV DNA in hybridization conditions of Denhardt solution, 2×

SSC, 50 mM sodium phosphate (pH 6.5), calf thymus DNA (250 μ

g/ml), and 35% deionized formamide at 42°

C., followed by washing with a wash solution of 60 mM Tris-hydrochloride solution (pH

     8), 2 mM EDTA, 300 mM NaCl, and 0.5% sodium dodecyl sulfate for 30 minutes at 50°

C., followed by 3 mM Tris solution for 30 minutes at room temperature.

6. A probe for the detection of human papillomavirus (HPV) DNA, which hybridizes under stringent conditions with the entire genome of a human papillomavirus selected from the group consisting of HPV-2d, HPV-10b, HPV-14a, HPV-14b, HPV-15, HPV-17a, HPV-17b, HPV-19, HPV-20, HPV-21, HPV-22, HPV-23, HPV-24, HPV-28, HPV-29, HPV-31, HPV-32, HPV-IP2 and HPV-IP4,

wherein said stringent conditions comprise contacting said probes with said HPV DNA in hybridization conditions of Denhardt solution, 2×

SSC, 50 mM sodium phosphate (pH 6.5), calf thymus DNA (250 μ

g/ml), and 35% deionized formamide at 42°

C., followed by washing with a wash solution of 60 mM Tris-hydrochloride solution (pH

     8), 2 mM EDTA, 300 mM NaCl, and 0.5% sodium dodecyl sulfate for 30 minutes at 50°

C., followed by 3 mM Tris solution for 30 minutes at room temperature.

7. A method for the in vitro detection of infection due to a papillomavirus, said method comprising

placing the probe of claim 6 in contact with nucleic acids of a biological sample under stringent hybridization conditions comprising hybridizing in Denhardt solution, 2×

SSC, 50 mM sodium phosphate (pH 6.5), calf thymus DNA (250 μ

g/ml), and 35% deionized formamide at 42°

C., followed by washing with a wash solution of 60 mM Tris-hydrochloride solution (pH

     8), 2 mM EDTA, 300 mM NaCl, and 0.5% sodium dodecyl sulfate for 30 minutes at 50°

C., followed by 3 mM Tris solution for 30 minutes at room temperature;

detecting a hybrid formed between DNA in the sample and said probe;

and correlating the presence of said hybrid with an infection due to the presence of a papillomavirus;

wherein said papillomavirus is selected from the group consisting of HPV-2d, HPV-10b, HPV-14a, HPV-14b, HPV-15, HPV-17a, HPV-17b, HPV-19, HPV-20, HPV-21, HPV-22, HPV-23, HPV-24, HPV-28, HPV-29, HPV-31, HPV-32, HPV-IP2 and HPV-IP4.

8. A kit comprising a plurality of different probes for detecting human papillomavirus (HPV) DNA, wherein said kit contains at least one probe selected from each of the following groups a-j:

a) DNA of HPV2d;

b) DNA of PV10b, 28, and 29;

c) DNA of HPV17 and 24;

d) DNA of HPV14, 15, 17, 19, 20, 21, 22 and 23;

e) DNA of HPV15 and 17;

f) DNA of HPV24;

g) DNA of HPV14 and 32;

h) DNA of HPV31;

i) DNA of HPV32; and

j) DNA of HPV16, 18 and HPV IP2;

wherein when said kit comprises one probe of each group, each of the said probes in said kit is of a different HPV type from the other probes.