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 detection reaction mixture comprising:
at least one detection probe comprising (a) a target-specific sequence consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO:13, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:52, complements thereof, and RNA equivalents thereof; and (b) a detectable label that is joined directly or indirectly to the target-specific sequence of (a).
2. The detection reaction mixture of claim 1 , wherein the detection reaction mixture further comprises a lithium salt selected from the group consisting of lithium succinate and lithium chloride.
3. The detection reaction mixture of claim 2 , wherein the reaction mixture comprises both lithium succinate and lithium chloride.
4. The detection reaction mixture of claim 1 , wherein the detection reaction mixture further comprises lithium lauryl sulfate.
5. The detection reaction mixture of claim 4 , wherein the detection reaction mixture further comprises a lithium salt selected from the group consisting of lithium succinate and lithium chloride.
6. The detection reaction mixture of claim 1 , wherein the detectable label is a chemiluminescent compound.
7. The detection reaction mixture of claim 6 , wherein the chemiluminescent compound is an acridinium ester (AE) compound.
8. An aqueous formulation comprising:
(a) at least one detection probe oligomer comprising (i) a target-specific sequence consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO:13, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:52, complements thereof, and RNA equivalents thereof, and (ii) a detectable label that is joined directly or indirectly to the target-specific sequence of (i);
(b) a lithium salt selected from the group consisting of lithium succinate and lithium chloride;
(c) lithium lauryl sulfate;
(d) EDTA; and
(e) ethanol.
9. The aqueous formulation of claim 8 , wherein the lithium salt is lithium succinate.
10. The aqueous formulation of claim 8 , wherein the lithium salt is lithium chloride.
11. The aqueous formulation of claim 8 , wherein the lithium lauryl sulfate is present in the formulation at a concentration of 2% (w/v).
12. The aqueous formulation of claim 11 , wherein the EDTA is present in the formulation at a concentration of 20 mM.
13. The aqueous formulation of claim 11 , wherein the formulation further comprises 15 mM mercaptoethanesulfonate.
14. The aqueous formulation of claim 8 , wherein the at least one detection probe oligomer comprises a target-specific sequence selected from the group consisting of SEQ ID NO:45, a complement thereof, and an RNA equivalent thereof.
15. The aqueous formulation of claim 8 , wherein the at least one detection probe oligomer comprises a target-specific sequence selected from the group consisting of SEQ ID NO:52, a complement thereof, and an RNA equivalent thereof.
16. An aqueous formulation comprising:
(a) one or more of (i) a detection probe oligomer comprising a target-specific sequence consisting of an RNA equivalent of SEQ ID NO:13, (ii) a detection probe oligomer comprising a target-specific sequence consisting of an RNA equivalent of SEQ ID NO:45, (iii) a detection probe oligomer comprising a target-specific sequence consisting of an RNA equivalent of SEQ ID NO:46, and (iv) a detection probe oligomer comprising a target-specific sequence consisting of an RNA equivalent of SEQ ID NO:52;
(b) 1.2 M lithium chloride;
(c) 2% (w/v) lithium lauryl sulfate;
(d) EDTA; and
(e) ethanol.
17. The aqueous formulation of claim 16 , further comprising 100 mM lithium succinate.
18. The aqueous formulation of claim 17 , further comprising 15 mM mercaptoethanesulfonate.
19. The aqueous formulation of claim 18 , wherein the EDTA is present in the formulation at a concentration of 20 mM.
20. The aqueous formulation of claim 16 , wherein at least one of the one or more detection probe oligomers of (i)-(vi) contains a 2′-methoxy backbone at one or more linkages in the nucleic acid backbone.