Digital assays with associated targets
AssigneeBio-Rad Laboratories, Inc.
InventorsAdam M. McCoy, Niels Klitgord
Life of this patent
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Abstract
System, including methods, apparatus, and compositions, for performing a digital assay with associated targets. In some embodiments, the assay of associated targets in partitions may lower the limit of detection (LOD) or otherwise increase assay sensitivity, accuracy, and/or specificity. The targets may represent spaced or overlapping regions of the same template and/or each may represent a region from a different complementary strand of the same template. In some embodiments, the associated targets may be provided by a type of biological particle, and the target content of partitions may be used to identify and quantify the type of biological particle in a sample.
Claims20
A method of performing a digital assay, the method comprising:
- forming partitions each including a portion of a sample that contains a first target associated with a second target;
- collecting data indicating a presence or absence of each target in individual partitions;
- classifying each of a plurality of the partitions as positive or negative for each of the targets based on the data, wherein partitions having an indicated presence of only the first target in the data are classified as negative for both targets;
- and
- determining a level of the first target.
A method of performing a multiplexed digital assay, the method comprising:
- providing partitions containing a single-stranded first template and a double-stranded second template having a first strand and a second strand, the first template and the second template each being present in only a subset of the partitions, the first template and the first strand each including a first target sequence and the second strand including a second target sequence;
- amplifying the first target sequence and the second target sequence in the partitions;
- collecting data for amplification of the first target sequence and the second target sequence in individual partitions;
- and
- determining a level of each of the single-stranded first template and the double-stranded second template based on the data.
A method of sample analysis, the method comprising:
- providing a sample comprising at least two types of biological particles and/or nucleic acid therefrom, wherein each type of biological particle contains a different target set or subset composed of at least one of two or more targets;
- forming partitions each containing a portion of the sample;
- determining a presence or absence of each of the two or more targets in each of a plurality of the partitions;
- and
- determining a level a type of biological particle containing a particular target set or subset of the two or more targets.
Dependencies are inferred from the claim text.