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US 2026/0092313 A1Published application
Nucleic acid detection methods
AssigneeThe Charles Stark Draper Laboratory, Inc.
InventorsIsaac Moran, Kim Kelly, Mike Turo, Erin Rosenberger
Life of this patent
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Abstract
The disclosure relates to methods and systems for the detection of low-quantity (e.g., 1×102 copies/μL) target nucleic acids (e.g., genomic RNA and/or genomic DNA).
Claims20
A method of detecting a target nucleic acid in a sample comprising:
- (i) contacting a sample in a reaction mixture with
- (a) a capture oligonucleotide that is immobilized to a surface via a silane-containing perfluorinated linker, wherein the capture oligonucleotide comprises a capture domain that is complementary to a first region of the target nucleic acid, and
- (b) a sense oligonucleotide that is functionalized with a first member of a binding pair and comprises a target-binding domain that is complementary to a second region of the target nucleic acid, wherein the first region of the target nucleic acid does not overlap with the second region of the target nucleic acid, and
- (ii) optionally removing nucleic acids that are not bound to the capture oligonucleotide from the reaction mixture,
- (iii) adding a detectable molecule that is conjugated to a second member of the binding pair to the reaction mixture, and
- (iv) detecting signal from surface-bound detectable label, wherein surface-bound signal at a level above background noise is indicative of the presence of the target nucleic acid in the sample.
3 more elements
A method of determining the concentration of a target nucleic acid in a sample, the method comprising:
- (i) contacting a sample comprising the target nucleic acid with a sense oligonucleotide and a capture oligonucleotide that is immobilized to a surface via a silane-containing perfluorinated linker to produce a surface-immobilized target nucleic acid, wherein the capture oligonucleotide comprises a capture domain that is complementary to a first region of the target nucleic acid, wherein the sense oligonucleotide is functionalized with a first member of a binding pair and comprises a target-binding domain that is complementary to a second region of the target nucleic acid, and wherein the first region of the target nucleic acid does not overlap with the second region of the target nucleic acid;
- (ii) contacting the surface-immobilized target nucleic acid with a detectable molecule that is conjugated to a second member of the binding pair;
- and
- (iii) determining the concentration of the target nucleic acid based on detection of the detectable molecule.
1 more element
A system for detecting a target nucleic acid in a sample, the system comprising:
- a substrate configured to receive a sample such that the sample is contacted in a reaction mixture with
- (a) a capture oligonucleotide that is immobilized to a surface on the substrate via a silane-containing perfluorinated linker, wherein the capture oligonucleotide comprises a capture domain that is complementary to a first region of the target nucleic acid, and
- (b) a sense oligonucleotide that is functionalized with a first member of a binding pair and comprises a target-binding domain that is complementary to a second region of the target nucleic acid, wherein the first region of the target nucleic acid does not overlap with the second region of the target nucleic acid,
- the substrate further configured to receive a detectable molecule that is conjugated to a second member of the binding pair such that the detectable molecule is added to the reaction mixture, and
- a detector configured to detect signal from surface-bound detectable label, wherein surface-bound signal at a level above background noise is indicative of the presence of the target nucleic acid in the sample.
2 more elements
A system for determining the concentration of a target nucleic acid in a sample, the system comprising:
- a substrate configured to receive the sample comprising the target nucleic acid such that the sample is contacted with a sense oligonucleotide and a capture oligonucleotide that is immobilized to a surface on the substrate via a silane-containing perfluorinated linker to produce a surface-immobilized target nucleic acid, wherein the capture oligonucleotide comprises a capture domain that is complementary to a first region of the target nucleic acid, wherein the sense oligonucleotide is functionalized with a first member of a binding pair and comprises a target-binding domain that is complementary to a second region of the target nucleic acid, and wherein the first region of the target nucleic acid does not overlap with the second region of the target nucleic acid,
- the substrate further configured to receive a detectable molecule that is conjugated to a second member of the binding pair such that the surface-immobilized target nucleic acid is contacted with the detectable molecule;
- and
- a processor configured to determine the concentration of the target nucleic acid based on detection of the detectable molecule.
1 more element
Dependencies are inferred from the claim text.