Detection of nucleic acid sequences by cleavage and separation of tag-containing structures
AssigneeMonogram Biosciences, Inc.
InventorsAhmed Chenna, Vivian Xiao, Sharat Singh
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Abstract
The present invention is directed to a method of detecting pluralities of target nucleic acid sequences by forming and cleaving duplex structures with a pair of probes, one probe of each pair being labeled with an electrophoretic tag. Cleavage of the duplex structures releases electrophoretic tags that are then separated and identified to indicate the presence or quantity of the target sequences. The present invention is particularly useful in multiplex reactions wherein multiple target sequences are detected in one reaction. Kits useful in the detection of nucleic acids are also provided.
Claims17
A method of detecting a plurality of polynucleotides in a sample, the method comprising the steps of:
- providing for each polynucleotide a helper probe complementary to a region of the polynucleotide and an electrophoretic probe complementary to the helper probe and to the polynucleotide adjacent to said region, such that the helper probe and the electrophoretic probe form a recognition duplex upon hybridization to each other and to the polynucleotide, each electrophoretic probe having attached an electrophoretic tag with a separation or detection characteristic distinct from those of other electrophoretic tags so that each electrophoretic tag forms a distinguishable peak in a separation profile;
- combining under hybridization conditions the sample, the helper probes, and the electrophoretic probes to form an assay mixture such that recognition duplexes are formed;
- cleaving the recognition duplexes at a cleavage site so that electrophoretic tags are released;
- and
- separating and identifying the released electrophoretic tags to detect each of the plurality of polynucleotides.
A kit for detecting the presence or absence of one or more target polynucleotides in a sample, the kit comprising:
- a plurality of pairs of helper probes and electrophoretic probes, each helper probe of a pair being complementary to a region of a target polynucleotide and each electrophoretic probe of the same pair being complementary to the helper probe and to the target polynucleotide adjacent to said region, such that the helper probe and the electrophoretic probe form a recognition duplex upon hybridization to each other and to the target polynucleotide, each electrophoretic probe having attached an electrophoretic tag with a separation or detection characteristic distinct from those of other electrophoretic tags so that each electrophoretic tag forms a distinguishable peak in a separation profile.
Dependencies are inferred from the claim text.