Detection of nucleic acids from multiple types of human papillomavirus
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.
1. A combination of oligomers for detecting human papillomavirus (HPV) types in group A2, the combination comprising:
a first amplification oligomer comprising a target-specific sequence and a 5′ promoter sequence, wherein the first amplification oligomer target-specific sequence consists of SEQ ID NO:25 or SEQ ID NO:27, or an RNA equivalent thereof; and
a second amplification oligomer comprising a target-specific sequence consisting of SEQ ID NO:38, or an RNA equivalent thereof.
2. The oligomer combination of claim 1 , wherein the nucleotide sequence of the first amplification oligomer consists of SEQ ID NO:24 or SEQ ID NO:26, or an RNA equivalent thereof.
3. The oligomer combination of claim 1 , wherein the first amplification oligomer target-specific sequence consists of SEQ ID NO:25, or an RNA equivalent thereof.
4. The oligomer combination of claim 3 , further comprising a third amplification oligomer comprising a target-specific sequence and a 5′ promoter sequence, wherein the third amplification oligomer target-specific sequence consists of SEQ ID NO:27, or an RNA equivalent thereof.
5. The oligomer combination of claim 4 , wherein
the nucleotide sequence of the first amplification oligomer consists of SEQ ID NO:24, or an RNA equivalent thereof, and
the nucleotide sequence of the third amplification oligomer consists of SEQ ID NO:26, or an RNA equivalent thereof.
6. The oligomer combination of claim 1 , further comprising a detection probe oligomer comprising a target-specific sequence selected from the group consisting of SEQ ID NO:12, a complement thereof, and an RNA equivalent thereof.
7. The oligomer combination of claim 6 , wherein the detection probe oligomer further comprises a detectable label.
8. The oligomer combination of claim 6 , wherein the detection probe oligomer target-specific sequence comprises at least one 2′-methoxy RNA group.
9. The oligomer combination of claim 1 , wherein the oligomer combination is contained in an amplification reaction mixture.
10. The oligomer combination of claim 6 , wherein the oligomer combination is contained in an amplified product detection reaction mixture.
11. The oligomer combination of claim 1 , wherein the oligomer combination is contained in a kit.
12. A method for of determining the presence or absence of a human papillomavirus (HPV) nucleic acid type in group A2 in a sample, comprising the steps of:
(a) contacting a sample, said sample suspected of containing a target nucleic acid of an HPV type in group A2, with a combination of amplification oligomers that amplify a target region of said target nucleic acid, wherein
(i) a first amplification oligomer comprises a target-specific sequence and a 5′ promoter sequence, wherein the first amplification oligomer target-specific sequence consists of SEQ ID NO:25 or SEQ ID NO:27, or an RNA equivalent thereof; and
(ii) a second amplification oligomer comprises a target-specific sequence consisting of SEQ ID NO:38, or an RNA equivalent thereof;
(b) performing an amplification reaction using the amplification oligomers, wherein if the target nucleic acid of the HPV type in group A2 is present in the sample, an amplification product is produced; and
(c) detecting the amplification product.
13. The method of claim 12 , wherein the first amplification oligomer target-specific sequence consists of SEQ ID NO:25, or an RNA equivalent thereof.
14. The method of claim 13 , wherein the combination of amplification oligomers further comprises a third amplification oligomer comprising a target-specific sequence and a 5′ promoter sequence, wherein the third amplification oligomer target-specific sequence consists of SEQ ID NO:27, or an RNA equivalent thereof.
15. The method of claim 12 , wherein the detecting step (c) comprises contacting the amplification product, if present, with a detection probe oligomer that binds specifically to the amplification product and results in a signal to indicate the presence in the sample of the HPV nucleic acid type in group A2.
16. The method of claim 15 , wherein the detection probe oligomer comprises a target-specific sequence selected from the group consisting of SEQ ID NO:12, a complement thereof, and an RNA equivalent thereof.
17. The method of claim 12 , further comprising a step of separating any target nucleic acid of the HPV type in group A2 from other components in the sample by (i) contacting the HPV target nucleic acid in the sample with a capture oligomer and (ii) separating a complex that includes the capture oligomer and the HPV target nucleic acid from other sample components before the amplifying step (b).
18. The method of claim 17 , wherein the capture oligomer comprises the target-specific sequence of SEQ ID NO:6 joined to a ligand moiety.
19. The method of claim 18 , wherein the ligand moiety is a substantially homopolymeric tail sequence attached to the 3′ end of the target-specific sequence of SEQ ID NO:6.
20. The method of claim 12 , wherein the amplification reaction of step (b) is a transcription-associated amplification reaction.