Kits for detecting HIV-2
View Patent ↗Compositions and methods for synthesizing and detecting HIV-2 specific amplicons. Particularly described are oligonucleotides that are useful as hybridization probes, and amplification primers that facilitate detection of very low levels of HIV-2 nucleic acids.
1. A kit for detecting HIV-2 nucleic acids, comprising a packaged combination of the following components in one or more containers:
(a) a first amplification oligonucleotide that consists of SEQ ID NO:10 and optionally comprises a 5′ promoter sequence non-complementary to the HIV-2 nucleic acid; and
(b) a second amplification oligonucleotide comprising a sequence of 19-40 contiguous bases from the sequence of SEQ ID NO:1, said second amplification oligonucleotide having a length of up to 100 nucleotides.
2. The kit of claim 1 , further comprising:
(c) an oligonucleotide detection probe that comprises the sequence of SEQ ID NO:21 or the complement thereof, and a detectable label.
3. The kit of claim 1 , wherein the length of the second amplification oligonucleotide is 19-40 nucleotides.
4. The kit of claim 3 , wherein the length of the first amplification oligonucleotide is up to 60 nucleotides.
5. The kit of claim 3 , wherein the length of the first amplification oligonucleotide is up to 60 nucleotides, wherein the first amplification oligonucleotide includes said optional 5′ promoter sequence.
6. The kit of claim 3 , wherein the length of the second amplification oligonucleotide is 19-21 nucleotides.
7. The kit of claim 4 , wherein the length of the second amplification oligonucleotide is 19-21 nucleotides.
8. The kit of claim 5 , wherein the first amplification oligonucleotide is the promoter-primer of SEQ ID NO:15.
9. The kit of claim 7 , wherein the second amplification oligonucleotide is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7.
10. The kit of claim 7 , wherein the first amplification oligonucleotide includes said optional 5′ promoter sequence.
11. The kit of claim 10 , wherein the first amplification oligonucleotide is the promoter-primer of SEQ ID NO:15.
12. The kit of claim 10 , wherein the second amplification oligonucleotide is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7.
13. The kit of claim 11 , wherein the second amplification oligonucleotide is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7.
14. The kit of claim 11 , wherein the length of the first amplification oligonucleotide is 25 nucleotides, and wherein the length of the second amplification oligonucleotide is 19-21 nucleotides.
15. The kit of claim 14 , wherein the second amplification oligonucleotide is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7.
16. The kit of claim 14 , wherein the second amplification oligonucleotide is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7.
17. The kit of claim 2 , wherein said oligonucleotide detection probe has a length of up to 18 nucleotides.
18. The kit of claim 17 , wherein the sequence of said oligonucleotide detection probe is selected from the group consisting of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26 and SEQ ID NO:27.
19. The kit of claim 18 , wherein the sequence of the first amplification oligonucleotide is SEQ ID NO:10 or SEQ ID NO:15, wherein the sequence of the second amplification oligonucleotide is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7, and wherein the sequence of the oligonucleotide detection probe is selected from the group consisting of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26 and SEQ ID NO:27.
20. The kit of claim 2 , wherein said oligonucleotide detection probe has a length of up to 35 nucleotides.
21. The kit of claim 20 , wherein the length of said oligonucleotide detection probe is up to 22 nucleotides.
22. The kit of claim 21 , wherein said oligonucleotide detection probe comprises at least 16 contiguous nucleotides contained within the sequence of SEQ ID NO:20 or the complement thereof.
23. The kit of claim 22 , wherein said oligonucleotide detection probe has the sequence of SEQ ID NO:20 or the complement thereof.
24. The kit of claim 22 , wherein said oligonucleotide detection probe has a length of up to 18 nucleotides.
25. The kit of claim 24 , wherein the length of said oligonucleotide detection probe is 18 nucleotides.
26. The kit of claim 24 , wherein said oligonucleotide detection probe has a sequence selected from the group consisting of SEQ ID NO:22 or the complement thereof, SEQ ID NO:23 or the complement thereof, SEQ ID NO:24 or the complement thereof, SEQ ID NO:25 or the complement thereof, SEQ ID NO:26 or the complement thereof, and SEQ ID NO:27 or the complement thereof.
27. The kit of claim 20 , wherein said oligonucleotide detection probe comprises DNA.
28. The kit of claim 20 , wherein said oligonucleotide detection probe comprises at least one nucleotide analog.
29. The kit of claim 28 , wherein said at least one nucleotide analog comprises a methoxy group at the 2′ position of a ribose moiety.
30. The kit of claim 26 , wherein the detectable label is a chemiluminescent label or a radiolabel.
31. The kit of claim 30 , wherein the detectable label is an acridinium ester.