Digital assays with multiplexed detection of two or more targets in the same optical channel
First Claim
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1. A method of performing a digital assay, comprising:
- amplifying at least two distinct targets in droplets containing a reporter for each distinct target, wherein the reporters each are labeled with a same luminophore;
detecting a signal from the droplets representing combined emission of light in a same wavelength regime from the reporters for the distinct targets, wherein each reporter provides a portion of the signal having an intensity that varies according to whether or not the corresponding distinct target is present in a droplet, and wherein a plurality of the droplets each contain at least two of the distinct targets and have a distinguishable intensity of the signal relative to droplets containing only one of the distinct targets;
determining for each distinct target a number of droplets that are positive for only one of the distinct targets;
determining at least one number of droplets that are positive for at least two of the distinct targets; and
determining a quantity representative of a level of at least one of the distinct targets.
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Abstract
System, including methods and apparatus, for performing a digital assay with multiplexed detection of two or more distinct targets in the same optical channel.
294 Citations
15 Claims
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1. A method of performing a digital assay, comprising:
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amplifying at least two distinct targets in droplets containing a reporter for each distinct target, wherein the reporters each are labeled with a same luminophore; detecting a signal from the droplets representing combined emission of light in a same wavelength regime from the reporters for the distinct targets, wherein each reporter provides a portion of the signal having an intensity that varies according to whether or not the corresponding distinct target is present in a droplet, and wherein a plurality of the droplets each contain at least two of the distinct targets and have a distinguishable intensity of the signal relative to droplets containing only one of the distinct targets; determining for each distinct target a number of droplets that are positive for only one of the distinct targets; determining at least one number of droplets that are positive for at least two of the distinct targets; and determining a quantity representative of a level of at least one of the distinct targets. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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Specification