Compositions for detecting BV-associated bacterial nucleic acid
Disclosed are nucleic acid oligomers, including amplification oligomers, capture probes, and detection probes, for detection of a 16S rRNA or its encoding gene from bacterial species associated with bacterial vaginosis. Also disclosed are methods of specific nucleic acid amplification and detection using the disclosed oligomers, as well as corresponding reaction mixtures and kits.
1. A reaction mixture for detection of a Megasphaera sp. target nucleic acid, said reaction mixture comprising:
(a) detection probe oligomer comprising
(i) a detection probe target-hybridizing sequence that consists of the sequence of SEQ ID NO:24; and
(ii) a chemiluminescent or fluorescent label; and
(b) an oligomer combination for amplification of a target region of the Megasphaera sp. target nucleic acid, wherein the target region contains the detection probe oligomer target sequence, said oligomer combination comprising
(i) a first amplification oligomer comprising a first target-hybridizing sequence that consists of the sequence of SEQ ID NO:15; and
(ii) a second amplification oligomer comprising a second target-hybridizing sequence that consists of the sequence of SEQ ID NO:21.
2. The reaction mixture of claim 1 , wherein the label is a chemiluminescent label.
3. The reaction mixture of claim 1 , wherein the label is a fluorescent label.
4. The reaction mixture of claim 3 , wherein the detection probe oligomer further comprises a quencher.
5. The reaction mixture of claim 4 , wherein the detection probe oligomer is a TaqMan detection probe or a molecular beacon.
6. The reaction mixture of claim 1 , wherein the detection probe oligomer further comprises a non-target-hybridizing sequence.
7. The reaction mixture of claim 6 , wherein the detection probe oligomer is a hairpin detection probe.
8. The reaction mixture of claim 7 , wherein the hairpin detection probe oligomer is a molecular beacon or a molecular torch.
9. The reaction mixture of claim 1 , wherein the second amplification oligomer is a promoter primer further comprising a promoter sequence located 5′ to the second target-hybridizing sequence.
10. The reaction mixture of claim 9 , wherein the promoter sequence is a T7 promoter sequence.
11. The reaction mixture of claim 9 , wherein the T7 promoter sequence consists of the sequence of SEQ ID NO:22.
12. The reaction mixture of claim 1 , further comprising a capture probe oligomer for purifying the Megasphaera sp. target nucleic acid.
13. The reaction mixture of claim 1 , further comprising a pseudotarget oligomer for detuning sensitivity of an assay for detecting the Megasphaera target nucleic acid.