Compositions, methods and kits to detect adenovirus nucleic acids
View Patent ↗The disclosed invention is related to methods, compositions, kits and isolated nucleic acid sequences for targeting Adenovirus nucleic acid. Compositions include amplification oligomers and/or detection probe oligomers. Kits and methods comprise at least one of these oligomers.
1. A method for detecting an Adenovirus nucleic acid comprising the steps of:
(a) contacting a test sample comprising nucleic acid with at least one hybridization assay probe having a nucleotide sequence consisting of a sequence selected from the group consisting of SEQ ID NOs: 10, 19, 21, 23, 24, 29, 36, and 37 or a combination of two or more thereof; and
(b) determining if a probe:target duplex has formed under stringent hybridization conditions in the test sample,
wherein the presence of the probe:target duplex is an indication of the presence of Adenovirus nucleic acid in the test sample.
2. The method of claim 1 , wherein the hybridization assay probe sequence consists of SEQ ID NO:10.
3. The method of claim 2 , wherein the contacting step further comprises contacting the test sample with a combination of at least two amplification oligomers wherein the combination comprises target hybridizing sequences of SEQ ID NOs:5 & 6, or SEQ ID NOs:5 & 8.
4. The method of claim 1 , wherein the hybridization assay probe comprises the sequence consisting of SEQ ID NO:19.
5. The method of claim 4 , wherein the contacting step further comprises contacting the test sample with a combination of at least two amplification oligomers wherein the combination comprises a first amplification oligomer comprising a target hybridizing sequence selected from SEQ ID NOs:11 & 12, and a second amplification oligomer comprising a target hybridizing sequence selected from SEQ ID NOs:13 & 15.
6. The method of claim 5 , wherein the hybridization assay probe is a Taqman probe comprising the sequence consisting of SEQ ID NO:19, a fluorophore attached to the sequence and a quencher attached to the sequence.
7. The method of claim 6 , wherein the first amplification oligomer comprises a target hybridizing sequences consisting of SEQ ID NO:12.
8. The method of claim 6 , wherein the second amplification oligomer comprises a target hybridizing sequences consisting of SEQ ID NO:15.
9. The method of claim 1 , wherein the at least one hybridization assay probe is a combination of two hybridization assay probes, one of which comprises a sequence consisting of SEQ ID NO:21 and the other of which comprises a sequence consisting of SEQ ID NO:24.
10. The method of claim 9 , wherein each of the two hybridization assay probes further comprises a fluorophore and a quencher attached to the sequence.
11. The method of claim 9 , wherein the contacting step further comprises contacting the test sample with a combination of at least two amplification oligomers wherein the combination comprises target hybridizing sequences of SEQ ID NOs:12 & 15.
12. The method of claim 1 , wherein the contacting step further comprises contacting the test sample with a combination of at least two amplification oligomers, and wherein the combination of at least two amplification oligomers and at least one hybridization assay probe comprises the sequences of:
(i) SEQ ID NOs: 21, 23, 25, 26, 27, & 28;
(ii) SEQ ID NOs:21, 23, 26, 27, & 28;
(iii) SEQ ID NOs:21, 23, 25, 27, & 28;
(iv) SEQ ID NOs:21, 23, 25, 26, & 28;
(v) SEQ ID NOs:21, 23, 25, 26, & 27;
(vi) SEQ ID NOs:23, 25, 26, 27, & 28;
(vii) SEQ ID NOs:25, 26, 27, 28, & 29;
(viii) SEQ ID NOs:21, 23, 27, 28, 33, & 34;
(ix) SEQ ID NOs:21, 23, 27, 28, 33, & 35;
(x) SEQ ID NOs:21, 23, 27, 28, 34, & 35; or
(xii) SEQ ID NOs:25, 26, 27, 28, 36, & 37.
13. The method of claim 12 , wherein each of the at least one hybridization assay probe sequences further comprises a fluorophore attached to the sequence.
14. The method of claim 13 , wherein each of the at least one hybridization assay probe sequences further comprises a quencher attached to the sequence.
15. A composition for use in detecting an Adenovirus target nucleic acid, wherein the composition comprises at least one hybridization assay probe comprising (A) a sequence selected from the group consisting of SEQ ID Nos. 10, 19, 21, 23, 24, 29, 36, and 37 or a combination of two or more thereof, and (B) a label attached to the sequence.
16. The composition of claim 15 , wherein the label is a fluorophore.
17. The composition of claim 16 , wherein the at least one hybridization assay probe further comprises a quencher that is attached to the sequence.