Encoded Nanopore Sensor for Multiplex Nucleic Acids Detection
AssigneeThe Curators of the University of Missouri
InventorsLi-Qun Gu, Xinyue Zhang
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Abstract
The present invention provides a new and improved multiplexed oligonucleotide detection method based on the nanopore technology with one or more probes containing a sequence with complementarity to the target oligonucleotide, a terminal extension at the probe's 3′ terminus, 5′ terminus, or both termini and a label attached to the terminus. The improved probes and probe sets enable sensitive, selective, and direct multiplex detection, differentiation and quantification of distinct target oligonucleotides such as miRNAs. The inventive detection method may also be employed as a non-invasive and cost-effective diagnostic method based on miRNA levels in the patient's tissue sample.
Claims21
1-52. (canceled)
A set of probe molecules including a first probe molecule and a second probe molecule, each comprising:
- a sequence partially complementary or fully complementary to a target nucleic acid;
- at least one extension covalently attached to the sequence;
- and
- at least one label covalently attached to the sequence or to an internal residue of the extension;
- wherein the sequence of the first probe molecule is partially complementary or fully complementary to a first target nucleic acid, and the sequence of the second probe molecule is partially complementary or fully complementary to a second target nucleic acid;
- and
- wherein the label of the first probe molecule is different from the label of the second probe molecule, such that the set of probe molecules provides distinct signals corresponding to the respective first target nucleic acid and second target nucleic acid in a nanopore system.
A set of probe molecules comprising a first probe molecule that comprises:
- a sequence partially complementary or fully complementary to a target nucleic acid;
- a first extension covalently attached to a 3′ terminus of the sequence, the first extension comprising a homopolymer, a di-nucleotide heteropolymer, or a tri-nucleotide heteropolymer;
- and
- a label covalently attached to an internal residue of the first extension within nine residues of the 3′ terminus of the sequence.
The set of probe molecules of claim 66, wherein the label comprises polyethylene glycol.
A set of probe molecules comprising:
- a first probe molecule comprising:
- a DNA sequence, an RNA sequence, or a peptide nucleic acid sequence partially complementary or fully complementary to a first target nucleic acid;
- and
- at least one label covalently attached to the sequence;
- and
- at least one second probe molecule comprising:
- a DNA sequence, an RNA sequence, or a peptide nucleic acid sequence partially complementary or fully complementary to a second target nucleic acid;
- wherein the set of probe molecules provides distinct signals corresponding to the respective first target nucleic acid and second target nucleic acid in a nanopore system.
Dependencies are inferred from the claim text.