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US 7,118,864 B2Granted
Amplifiable probe
AssigneeQuiatech AB
InventorMarek Kwiatkowski
Life of this patent
Estimating the term…
Abstract
Synthetic probes and methods for detecting analytes using such probes are described. The probes contain an analyte reactive moiety linked to a nucleic acid molecule that can be extended by DNA polymerase.
Claims18
A method of detecting the presence or absence of an analyte in a sample comprising:
- (a) contacting said sample with an amplifiable probe under conditions whereby said analyte, if present, binds said amplifiable probe to form a probe/analyte complex, said amplifiable probe comprising:
- (1) an analyte reactive moiety;
- (2) at least one extendable signal-generating nucleic acid, wherein each said at least one extendable signal-generating nucleic acid further comprises at least one self-complementary unit, wherein each said self-complementary unit comprises a first sequence element and a second sequence element, and wherein said first and said second nucleotide sequence elements are complementary;
- and
- (3) one or more linkers that connect said analyte reactive moiety to said at least one signal-generating nucleic acid;
- (b) extending said at least one extendable signal-generating nucleic acid in a reaction mixture comprising said probe/analyte complex, wherein said extendable signal-generating nucleic acid serves as its own template, wherein said extension comprises two or more cycles of extension, and wherein said extendable signal-generating nucleic acid is extended in each of said two or more cycles;
- and
- (c) detecting the presence or absence of said extended signal-generating nucleic acid, wherein presence of said extended signal-generating nucleic acid correlates with the presence of said analyte.
5 more elements
A method of detecting the presence or absence of an analyte in a sample comprising:
- (a) contacting said sample with an amplifiable probe under conditions whereby said analyte, if present, reacts with said amplifiable probe to form a modified amplifiable probe, said amplifiable probe comprising:
- (1) an analyte reactive moiety;
- (2) at least one extendable signal-generating nucleic acid, wherein each said at least one extendable signal-generating nucleic acid further comprises at least one self-complementary unit, wherein each said self-complementary unit comprises a first sequence element and a second sequence element, and wherein said first and said second sequence elements are complementary;
- and
- (3) one or more linkers that connect said analyte reactive moiety to said at least one signal-generating nucleic acid;
- (b) extending said at least one extendable signal-generating nucleic acid in a reaction mixture comprising said probe/analyte complex, wherein said extendable signal-generating nucleic acid serves as its own template, wherein said extension comprises two or more cycles of extension, and wherein said extendable signal-generating nucleic acid is extended in each of said two or more cycles;
- and
- (c) detecting the presence or absence of said extended signal-generating nucleic acid, wherein presence of said extended signal-generating nucleic acid correlates with the presence of said analyte.
5 more elements
A method of detecting the presence or absence of an analyte in a sample comprising:
- (a) contacting said sample with an amplifiable probe under conditions whereby said analyte, if present, binds said amplifiable probe to form a probe/analyte complex, said amplifiable probe comprising:
- (1) an analyte reactive moiety;
- (2) at least one signal-generating nucleic acid, wherein each said at least one signal-generating nucleic acid further comprises at least one self-complementary unit, wherein each said self-complementary unit comprises a first sequence element and a second sequence element, and wherein said first and said second nucleotide sequence elements are complementary;
- and
- (3) one or more linkers that connect said analyte reactive moiety to said at least one signal-generating nucleic acid;
- (b) contacting said probe/analyte complex with a cognate nucleic acid primer/template;
- (c) extending said cognate nucleic acid primer/template by performing an isothermal amplification reaction in a reaction mixture comprising said probe-analyte complex, wherein said extended cognate nucleic acid primer undergoes further self-primed extension;
- and
- (d) detecting the presence or absence of said extended signal-generating nucleic acid, wherein presence of said extended signal-generating nucleic acid correlates with the presence of said analyte.
6 more elements
Dependencies are inferred from the claim text.