Highly multiplexed particle-based assays
First Claim
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1. A method of detecting multiple analytes, which comprises:
- providing a first sample comprising at least one analyte from a first group of two or more analytes;
providing a second sample comprising at least one analyte from a second group of two or more analytes;
providing a first population of particles comprising two or more subsets of particles, wherein each subset of particles comprises a capture molecule that specifically binds directly or indirectly to one of the analytes of the first group, which capture molecule is different than that on all other subsets of particles in the first population, wherein a different capture molecule and subset of particles is provided for each different analyte in the first group;
providing a second population of particles comprising two or more subsets of particles, wherein each subset of particles comprises a capture molecule that specifically binds directly or indirectly to one of the analytes of the second group, which capture molecule is different than that on all other subsets of particles in the second population, wherein a different capture molecule and subset of particles is provided for each different analyte in the second group;
wherein each subset of particles is distinguishable from all other subsets of particles;
contacting the first sample with the first population of particles, and separately contacting the second sample with the second population of particles;
combining the first and second populations of particles after contacting them with the samples; and
after combining the populations of particles, detecting which subsets of particles have an analyte captured on the particles by detecting a label bound to the analyte, thereby indicating which analytes were present in the samples.
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Abstract
Methods are provided for detecting and optionally quantitating multiple analytes, including nucleic acid and/or polypeptide analytes, in particle-based assays that can be highly multiplexed. Compositions, systems, and kits related to the methods are also featured.
62 Citations
24 Claims
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1. A method of detecting multiple analytes, which comprises:
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providing a first sample comprising at least one analyte from a first group of two or more analytes; providing a second sample comprising at least one analyte from a second group of two or more analytes; providing a first population of particles comprising two or more subsets of particles, wherein each subset of particles comprises a capture molecule that specifically binds directly or indirectly to one of the analytes of the first group, which capture molecule is different than that on all other subsets of particles in the first population, wherein a different capture molecule and subset of particles is provided for each different analyte in the first group; providing a second population of particles comprising two or more subsets of particles, wherein each subset of particles comprises a capture molecule that specifically binds directly or indirectly to one of the analytes of the second group, which capture molecule is different than that on all other subsets of particles in the second population, wherein a different capture molecule and subset of particles is provided for each different analyte in the second group; wherein each subset of particles is distinguishable from all other subsets of particles; contacting the first sample with the first population of particles, and separately contacting the second sample with the second population of particles; combining the first and second populations of particles after contacting them with the samples; and after combining the populations of particles, detecting which subsets of particles have an analyte captured on the particles by detecting a label bound to the analyte, thereby indicating which analytes were present in the samples. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method of detecting multiple analytes, which comprises:
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providing a first sample comprising at least one analyte from a first group of five or more analytes; providing a second sample comprising at least one analyte from a second group of five or more analytes; providing a third sample comprising at least one analyte from a third group of five or more analytes; providing a first population of particles comprising five or more subsets of particles, wherein each subset of particles comprises a capture molecule that specifically binds directly or indirectly to one of the analytes of the first group, which capture molecule is different than that on all other subsets of particles in the first population, wherein a different capture molecule and subset of particles is provided for each different analyte in the first group; providing a second population of particles comprising five or more subsets of particles, wherein each subset of particles comprises a capture molecule that specifically binds directly or indirectly to one of the analytes of the second group, which capture molecule is different than that on all other subsets of particles in the second population, wherein a different capture molecule and subset of particles is provided for each different analyte in the second group; providing a third population of particles comprising five or more subsets of particles, wherein each subset of particles comprises a capture molecule that specifically binds directly or indirectly to one of the analytes of the third group, which capture molecule is different than that on all other subsets of particles in the third population, wherein a different capture molecule and subset of particles is provided for each different analyte in the third group; wherein each subset of particles is distinguishable from all other subsets of particles; separately contacting the first sample with the first population of particles, the second sample with the second population of particles, and the third sample with the third population of particles; combining the first, second, and third populations of particles after contacting them with the samples; and after combining the populations of particles, detecting which subsets of particles have an analyte captured on the particles by detecting a label bound to the analyte, thereby indicating which analytes were present in the samples. - View Dependent Claims (19, 20)
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21. A method of detecting multiple nucleic acid analytes, which comprises:
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providing a first sample comprising at least one nucleic acid analyte from a first group of two or more fluorescently labeled nucleic acid analytes; providing a second sample comprising at least one nucleic acid analyte from a second group of two or more fluorescently labeled nucleic acid analytes; providing a first population of particles comprising two or more subsets of particles, wherein each subset of particles comprises a DNA molecule that specifically binds directly or indirectly to one of the nucleic acid analytes of the first group, which DNA molecule is different than that on all other subsets of particles in the first population, wherein a different DNA molecule and subset of particles is provided for each different nucleic acid analyte in the first group; providing a second population of particles comprising two or more subsets of particles, wherein each subset of particles comprises a DNA molecule that specifically binds directly or indirectly to one of the nucleic acid analytes of the second group, which DNA molecule is different than that on all other subsets of particles in the second population, wherein a different DNA molecule and subset of particles is provided for each different nucleic acid analyte in the second group; wherein each subset of particles is distinguishable from all other subsets of particles by the color of the particles; contacting the first sample with the first population of particles, and separately contacting the second sample with the second population of particles; combining the first and second populations of particles after contacting them with the samples; and after combining the populations of particles, detecting which subsets of particles have a nucleic acid analyte captured on the particles, thereby indicating which nucleic acid analytes were present in the samples. - View Dependent Claims (22, 23, 24)
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Specification