METHODS OF DETECTING TARGETS ON AN ARRAY
First Claim
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1. A method of detecting a target analyte in a biological sample, comprising:
- a) providing a composite array comprising;
a substrate having a surface;
a first and a second assay location on said surface, wherein said assay locations comprise a population of microspheres, and wherein said microspheres comprise bioactive agents; and
a gasket separating said first assay location from said second assay location;
b) sealing said first and second assay locations thereby forming separately sealed hybridization chambers, said hybridization chambers each comprising a plurality of wells, wherein said microspheres are retained within said wells;
c) adding said biological sample to said first assay location under conditions sufficient to allow said target analyte to bind to said bioactive agents; and
d) detecting the binding of said bioactive agents to said target analyte, wherein the binding of said bioactive agents to said target analyte indicates the presence of said target analyte in said biological sample.
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Abstract
The invention relates to methods of detecting a target analyte in a biological sample using composite microsphere arrays having first and second assay locations. Preferred target analytes include nucleic acid, and more specifically, nucleic acid having one or more single nucleotide polymorphisms (SNPs).
111 Citations
20 Claims
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1. A method of detecting a target analyte in a biological sample, comprising:
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a) providing a composite array comprising; a substrate having a surface; a first and a second assay location on said surface, wherein said assay locations comprise a population of microspheres, and wherein said microspheres comprise bioactive agents; and a gasket separating said first assay location from said second assay location; b) sealing said first and second assay locations thereby forming separately sealed hybridization chambers, said hybridization chambers each comprising a plurality of wells, wherein said microspheres are retained within said wells; c) adding said biological sample to said first assay location under conditions sufficient to allow said target analyte to bind to said bioactive agents; and d) detecting the binding of said bioactive agents to said target analyte, wherein the binding of said bioactive agents to said target analyte indicates the presence of said target analyte in said biological sample. - View Dependent Claims (2, 3, 4)
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5. A method of detecting a target nucleic acid in a biological sample, wherein said target nucleic acid comprises one or more single nucleotide polymorphisms (SNPs) at one or more predetermined positions, comprising:
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a) providing a composite array comprising; a substrate having a surface; a population of microspheres, wherein said microspheres are linked to capture probes configured to bind to said target nucleic acid at said one or more predetermined positions; a first and second assay location on said surface, wherein said assay locations comprise said population of microspheres; and a gasket separating said first assay location from said second assay location; b) sealing said first and second assay locations thereby forming separately sealed hybridization chambers, said hybridization chambers each comprising a plurality of wells, wherein said microspheres are retained within said wells; c) adding said biological sample to said first assay location under conditions sufficient to allow said target nucleic acid to bind to said capture probes; and d) detecting the binding of said target nucleic acid to said capture probes, wherein the binding of said target analyte to said capture probes indicates the presence of said target nucleic acid in said biological sample. - View Dependent Claims (6, 7)
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8. A method of detecting a target analyte in a biological sample, comprising:
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a) providing a composite array comprising; a substrate having a surface; a first and a second assay location on said surface, wherein said assay locations comprise a population of microspheres, wherein said microspheres comprise bioactive agents and wherein said first and second assay locations each comprise a plurality of wells, wherein said microspheres are retained within said wells; a lid covering said first assay location and said second assay location; and a seal disposed between said lid and said surface, said seal separating said first assay location from said second assay location; b) adding said biological sample to said first assay location under conditions sufficient to allow said target analyte to bind to said bioactive agents; and c) detecting the binding of said bioactive agents to said target analyte, wherein the binding of said bioactive agents to said target analyte indicates the presence of said target analyte in said biological sample. - View Dependent Claims (9, 10, 11, 12, 13, 14)
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15. An apparatus, comprising:
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(a) composite array comprising a substrate with a surface comprising first and second assay locations separated from each other by a partition and a population of microspheres comprising a plurality of different bioactive agents, wherein said microspheres are randomly distributed on said surface; and (b) a detection instrument positioned to detect said microspheres. - View Dependent Claims (16, 17)
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18. An apparatus, comprising:
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(a) composite array comprising a substrate with a surface comprising first and second assay locations separated from each other by a partition and a population of microspheres comprising a plurality of subpopulations, wherein each subpopulation comprises a different bioactive agent, wherein said microspheres are randomly distributed on said surface; and (b) a fluid flow device attached to said composite array. - View Dependent Claims (19, 20)
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Specification