Light emission-or absorbance-based binding assays for polynucleic acids
AssigneeThe United States of America as represented by the Secretary of the Navy
InventorDavid A. Kidwell
Life of this patent
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Abstract
A target polynucleic acid strand may be detected within a sample. A sensing strand of complementary polynucleic acid is labelled, at at least two internucleotide phosphate groups, with labels which vary their electronically interaction with each other, and thus their emission or absorption spectra, depending upon whether the sensing strand is bound to target polynucleic acid in the sample.
Claims17
A method for detecting the presence of a target polynucleic acid strand having a specified nucleotide sequence in a matrix suspected of including said target polynucleic acid strand, comprising the steps of:
- adding to the matrix a strand of polynucleic acid complementary to said target polynucleic acid strand, said complementary polynucleic acid strand having attached thereto, at least two labels comprising fluorophore moieties, each attached to a corresponding internucleotide phosphate group, or between the phosphorus atoms of said complementary polynucleic acid, so that when said complementary polynucleic acid strand is not bound with said target polynucleic acid strand, said at least two fluorophore moieties are significantly more likely to electronically interact with each other to form an excimer and thus vary the wavelength dependence of their spectra than they would be if said complementary polynucleic acid strand were bound to said target polynucleic acid;
- and
- detecting said variation in the emission or absorption spectra, wherein the absence of, or a decrease in the intensity of, said variation in the emission or absorption spectra indicates the presence of said target polynucleic acid strand.
A method for detecting the presence of a target polynucleic acid strand having a specified nucleotide sequence in a matrix suspected of including said target polynucleic acid strand, comprising the steps of:
- adding to the matrix a strand of polynucleic acid complementary to said target polynucleic acid strand, said complementary polynucelic acid strand having attached thereto, at least two labels comprising fluorophore moieties, each attached to a corresponding internucleotide phosphate group and spaced sufficiently close together along said complementary strand so that when said complementary polynucelic acid is bound with said target polynucleic acid strand, said at least two fluorophore moieties are significantly more likely to electronically interact with each other to form an excimer and thus vary the wavelength dependance of their spectra than they would be if said complementary polynucleic acid strand were unbound to said target polynucleic acid strand;
- and
- detecting said variation in the emission or absorption spectra, wherein an increase in the intensity of said variation in the emission or absorption spectra indicates the presence of said target polynucleic acid strand.
Dependencies are inferred from the claim text.