Methods for detection and quantification of nucleic acid or protein targets in a sample

摘要

The invention provides an assay method for detection and/or quantification of a plurality of nucleic acid or protein targets in a sample. In the method probes are used to associate a detectable tag sequence with each of the selected targets present in the sample. Probes or primers sufficient to identify at least 25, and preferably at least 500, different targets are used. The method involves segregating aliquots of the sample from each other and detecting the tag sequences in each aliquot.

主权项

1. An assay method for detection of a plurality of nucleic acid or protein targets in a sample comprising
(a) combining target-specific probes with the sample under conditions in which at least 25 target-specific probes each bind a different specific target in the sample, if the specific target is present, wherein each said target-specific probe comprises a common nucleic acid tag sequence and a unique combination of a first primer binding site and a second primer binding site flanking the nucleic acid tag sequence, wherein at least two of said target-specific probes do not have the same first primer binding sites and at least two of said target-specific probes do not have the same second primer binding sites, and wherein the target-specific probe forms a target-specific probe product associated with the target if the target is present in the sample, thereby associating the tag with the target by forming a target-specific probe product associated with the target; (b) distributing aliquots of the sample to separate compartments of a microfluidic device and segregating the aliquots from each other; (c) querying each said aliquot for the presence or absence of a different target-specific probe product, wherein said querying comprises amplifying the nucleic acid tag sequence using a first primer that binds the first primer binding site and a second primer that binds the second primer binding site; and detecting the presence or absence of the common nucleic acid tag sequence using a tag-detecting probe.