Two stage nucleic acid amplification using an amplification oligomer
AssigneePanomics, Inc.
InventorsGary McMaster, Yunqing Ma
Life of this patent
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Abstract
This invention provides methods, compositions and systems to detect a nucleic acid of interest in a two-stage amplification. The two-stage amplification begins with a first non-enzymatic accumulation of an amplification oligomer that is the target substrate for a second nucleic acid amplification or assay. Two or more amplification oligomers can be used to allow multiplexed amplifications of two or more nucleic acids of interest with deconvolution based on unique detection signals or unique signal locations.
Claims29
1-31. (canceled)
An amplification oligomer comprising:
- a nucleotide sequence complimentary to an amplification multimer M-2 sequence of a first amplification multimer;
- and,
- a nucleotide sequence complimentary to a label extender L-2 sequence or to an M-1 sequence of a second amplification multimer.
46-59. (canceled)
A method of identifying amplification oligomers, the method comprising:
- providing a first label system comprising a first sequence complimentary to a first label probe comprising a first label;
- providing a second label system comprising a second sequence complimentary to a second label probe comprising a second label different from the first label;
- providing a sample comprising or suspected of comprising a first amplification oligomer comprising a sequence complimentary to a component of the first label system, or which sample comprises or is suspected of comprising a second amplification oligomer comprising a sequence complimentary to a component of the second label system;
- hybridizing the sample with the first and second label system;
- and,
- detecting a signal from the first label or from the second label;
- wherein detection of the signal from the first label indicates the presence of the first amplification oligomer in the sample or detection of the signal from the second label indicates the presence of the second amplification oligomer in the sample.
A method of identifying a nucleic acid of interest in a sample, which method comprises:
- providing a sample comprising or suspected of comprising one or more nucleic acids of interest;
- providing a first solid support which comprises a first capture probe sequence or a first capture extender sequence complimentary to a first nucleic acid of interest;
- providing a second solid support which comprises a second capture probe sequence or a second capture extender sequence complimentary to a second nucleic acid of interest;
- contacting the first and second solid supports with the sample;
- capturing the first nucleic acid of interest on the first solid support or capturing the second nucleic acid of interest on the second solid support;
- hybridizing a first labeling system directly or indirectly to the first or second nucleic acid of interest;
- hybridizing the first labeling system to one or more amplification oligomers;
- separating the first solid support from the second solid support;
- and,
- separately detecting the amplification oligomers that have been hybridized on each solid support in a nucleic acid assay;
- whereby the presence of the first nucleic acid of interest is identified if the amplification oligomers are detected on the first solid support and presence of the second nucleic acid of interest is identified if the amplification oligomers are detected on the second solid support.
Dependencies are inferred from the claim text.