Devices and methods for detecting nucleic acid analytes in samples
First Claim
1. An assay device for detecting the presence of a nucleic acid analyte in a sample, said device comprising a planar, solid support having covalently bound thereto a two-dimensional field of analyte-specific nucleic acid probes, each probe comprising a first end covalently bound to said support and including a 5'"'"' portion of an autocatalytically replicable nucleic acid, and a second end including an analyte-binding segment.
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Accused Products
Abstract
The invention provides devices and methods for use in detecting nucleic acid analytes in samples. The devices each include a solid support to which is bound a two-dimensional distribution or field of nucleic acid probes that each bind to a nucleic acid analyte, which is detected by use of amplification methods.
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Citations
25 Claims
- 1. An assay device for detecting the presence of a nucleic acid analyte in a sample, said device comprising a planar, solid support having covalently bound thereto a two-dimensional field of analyte-specific nucleic acid probes, each probe comprising a first end covalently bound to said support and including a 5'"'"' portion of an autocatalytically replicable nucleic acid, and a second end including an analyte-binding segment.
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5. A method for detecting the presence of a nucleic acid analyte in a sample, said method comprising the steps of:
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(a) applying the sample to a solid support having covalently bound thereto a two-dimensional field of first analyte-specific nucleic acid probes, each of the first probes comprising a first end bound to the solid support and including a 5'"'"' portion of an autocatalytically replicable nucleic acid, and a second end including a first analyte-binding segment, wherein the first analyte-binding segment of each of the first probes hybridizes to a first region of the analyte; (b) applying a second nucleic acid probe to the solid support, the second probe comprising a second analyte-binding segment that hybridizes to a second region of the analyte, the second probe further comprising a remainder of said autocatalytically replicable nucleic acid, wherein the first and second regions of the analyte are adjacent nucleotide segments, and wherein the analyte, the first probe, and the second probe hybridize together on the support to form a complex including a complete autocatalytically replicable nucleic acid; (c) applying a diffusion limiting matrix to the solid support; (d) amplifying the complete autocatalytically replicable nucleic acid to generate an amplified product; and (e) detecting the amplified product as a measure of the presence of the analyte in the sample. - View Dependent Claims (6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. An assay system for detecting a nucleic acid analyte in a sample, said assay system comprising:
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(a) a planar solid support having covalently bound thereto a two-dimensional field of first analyte-specific nucleic acid probes, each of the first probes comprising a first end covalently bound to the solid support and including a 5'"'"' portion of an autocatalytically replicable nucleic acid, and a second end including a first analyte-binding segment, wherein the first analyte-binding segment of each of the first probes hybridizes to a first region of the analyte; and (b) a second nucleic acid probe comprising a second analyte-binding segment that hybridizes to a second region of the analyte, the second probe further comprising a remainder of the autocatalytically replicable nucleic acid, and the first and second regions of the analyte comprising adjacent nucleotide segments; wherein binding of the first and second probes to the analyte permits amplification of the autocatalytically replicable nucleic acid.
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19. An assay system for detecting a nucleic acid analyte in a sample, said assay system comprising:
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(a) a solid support having bound thereto a two-dimensional field of capture probes, each of the capture probes comprising a capture segment that hybridizes to a capture region of the analyte; (b) a first nucleic acid probe comprising a first segment that hybridizes to a first region of the analyte; (c) a second nucleic acid probe comprising a second segment that hybridizes to a second region of the analyte; and (d) a diffusion limiting matrix for application to the solid support; wherein binding of the capture probes and the first and second probes to the analyte permits amplification of a detectable product within the diffusion limiting matrix. - View Dependent Claims (20, 21, 22, 23)
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24. A method for detecting the presence of a nucleic acid analyte in a sample, the method comprising the steps of:
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(a) contacting the sample with a solid support having covalently bound thereto a two-dimensional field of first analyte-specific nucleic acid probes, each of the first probes comprising a first end bound to the support and including a second end including a first analyte-binding segment; (b) applying a diffusion limiting matrix to the support; (c) performing amplification by one of polymerase chain reaction, ligase chain reaction, transcription-mediated amplification, nucleic acid sequence-based amplification, and strand displacement amplification in a manner dependent upon the presence of both the first probe and the nucleic acid analyte, and in such a manner that the products of such amplification reaction are restrained to form localized foci within the diffusion limiting matrix on said support; and (d) detecting the amplified products as a measure of the presence of the analyte in the sample.
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25. An assay system for detecting a nucleic acid analyte in a sample, said assay system comprising:
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(a) a solid support having covalently bound thereto a two-dimensional field of first analyte-specific nucleic acid probes, each of the first probes comprising a first end covalently bound to the solid support and including a 5'"'"' portion of an autocatalytically replicable nucleic acid, and a second end including a first analyte-binding segment, wherein the first analyte-binding segment of each of the first probes hybridizes to a first region of the analyte; (b) a second nucleic acid probe comprising a second analyte-binding segment that hybridizes to a second region of the analyte, the second probe further comprising a remainder of the autocatalytically replicable nucleic acid, and the first and second regions of the analyte comprising adjacent nucleotide segments; and (c) a diffusion limiting matrix for application to the solid support; wherein binding of the first and second probes to the analyte permits amplification of the autocatalytically replicable nucleic acid within the diffusion limiting matrix.
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Specification