Compositions, reaction mixtures and methods for detecting nucleic acids from type A1 and/or type C1 human papillomavirus

• US 8,574,841 B2
• Filed: 11/15/2012
• Issued: 11/05/2013
• Est. Priority Date: 12/08/2004
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: Norman, Sylvia A., Bungo, Jennifer J., Hanna, William L., Rao, Neeraj P.
• Primary Examiner(s): Babic, Christopher M.
• Application Number:
  US13/678,402
• Publication Number:
  US 20130078618A1
• Time in Patent Office: 355 Days
• Field of Search: None
• US Class Current: 435/6.1
• 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: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

Nucleic acid oligonucleotide sequences are disclosed which include amplification oligomers and probe oligomers which are useful for detecting multiple types of human papillomaviruses (HPV) associated with cervical cancer. Methods for detecting multiple HPV types in biological specimens by amplifying HPV nucleic acid sequences in vitro and detecting the amplified products are disclosed.

Claims

1. A kit containing at least two detection probes specific for a type A1 or type C1 HPV target region, wherein each of the detection probes individually has a contiguous base sequence that is 20 to 28 nucleotides in length and is at least 90% identical to a sequence selected from the group consisting of:

SEQ ID NOs;

11, 13, 44-47, 49, 51, 52, and 54, complements, RNA equivalents, and RNA/DNA chimerics thereof,

wherein at least one detection probe is specific for a type A1 HPV nucleic acid and at least one detection probe is specific for a type C1 HPV nucleic acid, and

wherein the detection probe specific for a type A1 HPV nucleic acid is joined to an AE label that is different than an AE label to which the detection probe specific for a type C1 HPV nucleic acid is joined, in order to provide distinguishable signals.

2. The kit of claim 1, wherein the kit further contains components for making a Probe Reagent, components for making a Selection Reagent, components for making a Detection Reagent, or combinations thereof.

3. The kit of claim 1, wherein the kit further contains at least one amplification oligonucleotide selected from the group consisting of SEQ ID NOS:

18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 38, 39 and 42.

6. The kit of claim 1, wherein each of the at least two detection probes individually has a contiguous base sequence selected from the group consisting of:

SEQ ID NOs;

11, 13, 44-47, 49, 51, 52, and 54, complements, RNA equivalents, and RNA/DNA chimerics thereof.

7. The kit of claim 1, wherein each of the at least two detection probes contains a 2′

-methoxy backbone at one or more linkages in the nucleic acid backbone.

4. A detection reaction mixture for detecting the presence of a type A1 HPV nucleic acid and/or a type C1 HPV nucleic acid in a sample, wherein the reaction mixture contains at least two detection probes specific for a type A1 or type C1 HPV target region, wherein each of the at least two detection probes individually has a contiguous base sequence that is 20 to 28 nucleotides in length and is at least 90% identical to a sequence selected from the group consisting of:

SEQ ID NOs;

11, 13, 44-47, 49, 51, 52, and 54, complements, RNA equivalents, and RNA/DNA chimerics thereof,

wherein at least one detection probe is specific for a type A1 HPV nucleic acid and at least one detection probe is specific for a type C1 HPV nucleic acid, and

wherein the detection probe specific for a type A1 HPV nucleic acid is joined to an AE label that is different than an AE label to which the detection probe specific for a type C1 HPV nucleic acid is joined, in order to provide distinguishable signals.

8. The detection reaction mixture of claim 4, wherein each of the at least two detection probes individually has a contiguous base sequence selected from the group consisting of:

SEQ ID NOs;

11, 13, 44-47, 49, 51, 52, and 54, complements, RNA equivalents, and RNA/DNA chimerics thereof.

9. The detection reaction mixture of claim 4, wherein each of the at least two detection probes contains a 2′

-methoxy backbone at one or more linkages in the nucleic acid backbone.

5. A method for detecting the presence of a human papillomavirus (HPV) nucleic acid in a sample comprising the steps of:

(a) treating a biological sample suspected of containing a type A1 HPV nucleic acid and/or a type C1 HPV nucleic acid to release intracellular components;

(b) separating an HPV nucleic acid away from a component of the biological sample;

(c) contacting the separated HPV nucleic acid with at least two detection probes specific for a type A1 or type C1 HPV target region, wherein each of the at least two detection probes individually has a contiguous base sequence that is 20 to 28 nucleotides in length and is at least 90% identical to a sequence selected from the group consisting of;

SEQ ID NOs;

11, 13, 44-47, 49, 51, 52, and 54, complements, RNA equivalents, and RNA/DNA chimerics thereof

wherein at least one detection probe is specific for a type A1 HPV nucleic acid and at least one detection probe is specific for a type C1 HPV nucleic acid, and

wherein the detection probe specific for a type A1 HPV nucleic acid is joined to an AE label that is different than an AE label to which the detection probe specific for a type C1 HPV nucleic acid is joined, in order to provide distinguishable signals; and

(d) detecting a signal that results from hybridization of at least one of the detection probes to an HPV target region to indicate the presence of a type A1 and/or type C1 HPV nucleic acid in the sample.

10. The method of claim 5, wherein each of the at least two detection probes contains a 2′

-methoxy backbone at one or more linkages in the nucleic acid backbone.

11. The method of claim 5, wherein the separated HPV nucleic acid from step (b) is amplified in an in vitro amplification reaction that includes a combination of amplification oligonucleotides that amplify a HPV type A1 target sequence and a combination of amplification oligomers that amplify HPV type C1 target sequence in an E6/E7 target region sequence.

12. The method of claim 11, wherein the combination of amplification oligonucleotides that amplify a HPV type A1 target sequence includes at least one amplification oligonucleotide selected from the group consisting of:

SEQ ID NOS;

18, 19, 20, 21, 22, 23, 38 and 42, complements, RNA equivalents, and RNA/DNA chimerics thereof.

13. The method of claim 11, wherein the combination of amplification oligonucleotides that amplify a HPV type C1 target sequence includes at least one amplification oligonucleotide selected from the group consisting of:

SEQ ID NOS;

28, 29, 30, 31 and 39, complements, RNA equivalents, and RNA/DNA chimerics thereof.

14. The method of claim 11, wherein each of the at least two detection probes individually has a contiguous base sequence selected from the group consisting of:

SEQ ID NOs;

11, 13, 44-47, 49, 51, 52, and 54, complements, RNA equivalents, and RNA/DNA chimerics thereof.

15. The method of claim 14, wherein (i) the combination of amplification oligonucleotides that amplify a HPV type A1 target sequence includes at least one amplification oligonucleotide that has a contiguous base sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOS:

18, 19, 20, 21, 22, 23, 38 and 42, complements, RNA equivalents, and RNA/DNA chimerics thereof, and wherein (ii) the combination of amplification oligonucleotides that amplify a HPV type C1 target sequence includes at least one amplification oligonucleotide that has a contiguous base sequence that is at least 90% identical to a sequence selected from the group consisting of;

SEQ ID NOS;

28, 29, 30, 31 and 39, complements, RNA equivalents, and RNA/DNA chimerics thereof.

16. The method of claim 11, wherein the in vitro amplification reaction occurs before step (c).

17. The method of claim 5, wherein the separating step (b) uses a capture oligonucleotide, wherein a capture oligonucleotide is selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2 joined to a ligand moiety, SEQ ID NO;

5, SEQ ID NO;

6 joined to a ligand moiety, and combinations thereof.

18. The method of claim 5, wherein the biological sample is treated to physically disrupt the sample and release intracellular components.

19. The method of claim 5, wherein the biological sample is treated with an enzyme to release intracellular components.

20. The method of claim 5, wherein the separating step (b) is a target capture step that uses a target capture reaction mixture containing, (i) a capture oligonucleotide selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2 joined to a ligand moiety, SEQ ID NO;

5, SEQ ID NO;

6 joined to a ligand moiety, and combinations thereof, and (ii) at least one amplification oligonucleotide selected from the group consisting of SEQ ID NOS;

18, 19, 20, 21, 22, 23, 28, 29, 30, 31, 38, 39 and 42.