AMPLIFICATION OF NUCLEIC ACIDS WITH ELECTRONIC DETECTION
First Claim
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1. A method for detecting a first target nucleic acid sequence comprising:
- a) hybridizing at least a first primer nucleic acid to said first target sequence to form a first hybridization complex;
b) contacting said first hybridization complex with a first enzyme to form a modified first primer nucleic acid;
c) disassociating said first hybridization complex;
d) forming a first assay complex comprising at least one ETM and said modified first primer nucleic acid, wherein said first assay complex is covalently attached to an electrode; and
e) detecting electron transfer between said ETM and said electrode as an indication of the presence of said target sequence.
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Abstract
The invention relates to compositions and methods useful in the detection of nucleic acids using a variety of amplification techniques, including both signal amplification and target amplification. Detection proceeds through the use of an electron transfer moiety (ETM) that is associated with the nucleic acid, either directly or indirectly, to allow electronic detection of the ETM using an electrode.
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Citations
20 Claims
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1. A method for detecting a first target nucleic acid sequence comprising:
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a) hybridizing at least a first primer nucleic acid to said first target sequence to form a first hybridization complex;
b) contacting said first hybridization complex with a first enzyme to form a modified first primer nucleic acid;
c) disassociating said first hybridization complex;
d) forming a first assay complex comprising at least one ETM and said modified first primer nucleic acid, wherein said first assay complex is covalently attached to an electrode; and
e) detecting electron transfer between said ETM and said electrode as an indication of the presence of said target sequence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A method for detecting a target nucleic acid sequence comprising:
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a) forming a first hybridization complex comprising an amplifier probe and a target sequence, wherein said amplifier probe comprises at least two amplification sequences;
b) hybridizing a first portion of at least one label probe to all or part of at least one amplification sequence;
c) hybridizing a second portion of said label probe to a detection probe covalently attached to an electrode via a conductive oligomer to form a second hybridization complex that contains at least a first electron transfer moiety (ETM); and
d) detecting said label probe by measuring electron transfer between said first ETM and said electrode.
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14. A method for detecting a target nucleic acid sequence comprising:
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a) forming a first hybridization complex comprising an amplifier probe and a target sequence, wherein said amplifier probe comprises at least two amplification sequences, wherein said first hybridization complex is covalently attached to an electrode comprising a monolayer comprising conductive oligomers;
b) hybridizing at least one label probe comprising at least one electron transfer moiety (ETM) to all or part of at least one amplification sequence;
c) detecting said label probe by measuring electron transfer between said first ETM and said electrode.
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15. A kit for the detection of a first target nucleic acid sequence comprising:
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a) at least a first nucleic acid primer substantially complementary to at least a first domain of said target sequence;
b) at least a first enzyme that will modify said first nucleic acid primer; and
c) an electrode comprising at least one detection probe covalently attached to said electrode via a conductive oligomer. - View Dependent Claims (17, 18, 19, 20)
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16. A kit for the detection of a first target nucleic acid sequence comprising:
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a) at least a first nucleic acid primer substantially complementary to at least a first domain of said target sequence;
b) at least a first enzyme that will modify said first nucleic acid primer; and
c) an electrode comprising a monolayer comprising conductive oligomers.
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Specification