COMPOSITIONS TO DETECT ADENOVIRUS NUCLEIC ACIDS
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 composition comprising a detection probe configured to hybridize to a portion of an Adenovirus nucleic acid or to a portion of an amplicon generated therefrom, wherein the detection probe comprises
(a) a target hybridizing sequence selected from the group consisting of SEQ ID NO:10, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:39 and SEQ ID NO:40; and
(b) one or more of
(i) a label,
(ii) a substitution at the 2′ position of at least one ribose moiety, and
(iii) a blocking moiety at or near the 3′ end of the detection probe, wherein the blocking moiety is configured to prevent enzyme-mediated extension of the detection probe in an amplification reaction.
2 . The composition of claim 1 , wherein the detection probe comprises the label and the label is selected from the group consisting of a fluorescent label, a chemiluminescent label, a radiolabel, an enzyme label, a hapten label, a linked oligonucleotide label, or an electronically detectable label.
3 . The composition of claim 1 , wherein the detection probe comprises an interactive pair of labels.
4 . The composition of claim 3 , wherein the interactive pair of labels is a fluorescent label in combination with a quencher.
5 . The composition of claim 3 , wherein the detection probe is a hairpin probe.
6 . The composition of claim 5 , wherein the hairpin probe is a molecular torch or a molecular beacon.
7 . A composition comprising at least two detection probes configured to hybridize to a portion of an Adenovirus nucleic acid or to a portion of an amplicon generated therefrom, wherein each of the detection probes comprises
(a) a target hybridizing sequence selected from the group consisting of SEQ ID NO:10, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:39 and SEQ ID NO:40; and
(b) one or more of
(i) a label,
(ii) a substitution at the 2′ position of at least one ribose moiety, and
(iii) a blocking moiety at or near the 3′ end of the detection probe, wherein the blocking moiety is configured to prevent enzyme-mediated extension of the detection probe in an amplification reaction.
8 . The composition of claim 7 , wherein each of the at least two detection probes comprises the label and the label is selected from the group consisting of a fluorescent label, a chemiluminescent label, a radiolabel, an enzyme label, a hapten label, a linked oligonucleotide label, or an electronically detectable label.
9 . The composition of claim 7 , wherein each of the at least two detection probes comprises an interactive pair of labels.
10 . The composition of claim 9 , wherein the interactive pair of labels is a fluorescent label in combination with a quencher.
11 . The composition of claim 9 , wherein each of the at least two detection probes is a hairpin probe.
12 . The composition of claim 11 , wherein the hairpin probe is a molecular torch or a molecular beacon.
13 . A hybridization assay reaction mixture comprising:
(1) a detection probe configured to hybridize to a portion of an Adenovirus nucleic acid or to a portion of an amplicon generated therefrom, wherein the detection probe comprises
(a) a target hybridizing sequence selected from the group consisting of SEQ ID NO:10, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:39 and SEQ ID NO:40; and
(b) one or more of
(i) a label,
(ii) a substitution at the 2′ position of at least one ribose moiety, and
(iii) a blocking moiety at or near the 3′ end of the detection probe, wherein the blocking moiety is configured to prevent enzyme-mediated extension of the detection probe in an amplification reaction; and
(2) a salt.
14 . The reaction mixture of claim 13 , wherein the detection probe comprises the label and the label is a homogeneous detectable label.
15 . The reaction mixture of claim 14 , wherein the homogeneous detectable label is a fluorescent label, a chemiluminescent label, or an electronically detectable label.
16 . The reaction mixture of claim 13 , wherein the detection probe comprises an interactive pair of labels.
17 . The reaction mixture of claim 16 , wherein the interactive pair of labels is a fluorescent label in combination with a quencher.
18 . The reaction mixture of claim 16 , wherein the detection probe is a hairpin probe.
19 . The reaction mixture of claim 18 , wherein the hairpin probe is a molecular torch or a molecular beacon.
20 . The reaction mixture of claim 13 , wherein the salt is selected from the group consisting of lithium chloride, sodium chloride, and sodium citrate.