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US5332659A Expired / Lapsed US ⏱️ 0.0 years remaining

Light emission-or absorbance-based binding assays for polynucleic acids

Assignee: United States Department of the Navy
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Filing: 1993-01-15
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Granted: 1994-07-26
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Expires: 2013-01-15
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Abstract

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.

Inventors

  • 👤 David A. Kidwell

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-29 20:03
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6818; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/542; G01N33/94; G01N33/946

Claims Analysis

Total Claims: 19
Claim 1
Independent Claim 182 words
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.
Claim 2
Independent Claim 182 words
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.
Claim 3
Dependent Claim 17 words
The method of claim 1, wherein said variation occurs in the emission spectra of said fluorophore moieties.
Claim 4
Dependent Claim 17 words
The method of claim 1, wherein said variation occurs in the absorption spectra of said fluorophore moieties.
Claim 5
Dependent Claim 24 words
The method of claim 1, wherein each of said at least two labels is attached to its corresponding internucleotide phosphate group by a linker.
Claim 6
Dependent Claim 21 words
The method of claim 4, wherein the linker is soluble in said matrix and is between 1 and 1300 atoms long.
Claim 7
Dependent Claim 17 words
The method of claim 1 wherein each of said at least two labels is a fluorescent dye.
Claim 8
Dependent Claim 18 words
The method of claim 6, wherein each of said at least two labels is a polycyclic aromatic compound.
Claim 9
Dependent Claim 24 words
The method of claim 7, wherein each of said at least two labels includes a pyrene ring, a napthalene ring or an anthracene ring.
Claim 10
Dependent Claim 11 words
The method of claim 1, wherein the matrix includes a surfactant.
Claim 11
Independent Claim 176 words
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.
Claim 12
Independent Claim 176 words
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.
Claim 13
Dependent Claim 17 words
The method of claim 10, wherein said variation occurs in the emission spectra of said fluorophore moieties.
Claim 14
Dependent Claim 17 words
The method of claim 10, wherein said variation occurs in the absorption spectra of said fluorophore moieties.
Claim 15
Dependent Claim 24 words
The method of claim 10, wherein each of said at least two labels is attached to its corresponding internucleotide phosphate group by a linker.
Claim 16
Dependent Claim 21 words
The method of claim 13, wherein the linker is soluble in said matrix and is between 1 and 1300 atoms long.
Claim 17
Dependent Claim 17 words
The method of claim 10, wherein each of said at least two labels is a fluroescent dye.
Claim 18
Dependent Claim 18 words
The method of claim 15, wherein each of said at least two labels is a polycyclic aromatic compound.
Claim 19
Dependent Claim 24 words
The method of claim 13, wherein each of said at least two labels includes a pyrene ring, a napthalene ring or an anthracene ring.

References Cited (Backward Citations)

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USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1997-07-25 1998-01-24 1998-07-26 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 2001-07-25 2002-01-24 2002-07-26 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2005-07-25 2006-01-24 2006-07-26 $7700.00 Window Closed / Lapsed

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