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US 2003/0165935 A1Published application
Digital assay
OwnerCharles S. Vann, Kai Q. Laoinventors
Life of this patent
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Abstract
The invention relates to methods and compositions for the detection of targets in a sample.
Claims38
A method for quantitating a target comprising;
- forming a reaction mixture comprising:
- a sample possibly containing the target;
- a codeable label;
- one or more target-specific probes, wherein each target-specific probe binds specifically to the target under selective binding conditions;
- and a separating moiety;
- treating the reaction mixture under reaction conditions such that a detectable complex is produced when the target is present, and wherein the detectable complex comprises the codeable label, the target-specific probe, and the separating moiety;
- and
- quantitating the target by counting the number of codeable labels.
5 more elements
A method for quantitating at least two different particular targets comprising;
- forming a reaction mixture comprising:
- a sample possibly containing two or more different particular targets;
- a different codeable label specific for each different particular target;
- one or more different target-specific probes specific for each different particular target that bind specifically to the target under selective binding conditions;
- and a separating moiety;
- treating the reaction mixture under reaction conditions such that when a particular target is present, a detectable complex is produced, which comprises the codeable label specific for the particular target, the target-specific probe specific for the particular target, and the separating moiety;
- and
- quantitating each of the different particular targets by counting the number of codeable labels specific for each of the different particular targets.
5 more elements
A method for quantitating at least two different target nucleic acid sequences in a sample comprising:
- forming a ligation reaction mixture by combining the sample with a different probe set specific for each of the at least two different target nucleic acid sequences, each probe set comprising (a) at least one separating bead, comprising a magnetic particle and a first target-specific probe, and (b) at least one detecting bead, comprising a codeable label, and a second target-specific probe;
- wherein the target-specific probes in each set are suitable for ligation together when hybridized adjacent to one another on a complementary target sequence;
- subjecting the ligation reaction mixture to a ligation reaction, wherein adjacently hybridizing complementary target-specific probes are ligated to one another to form a ligation product comprising the separating bead and the detecting bead;
- and
- quantitating each of the at least two different target nucleic acid sequences by counting the number of codeable labels.
2 more elements
A kit for detecting target nucleic acid sequences in a sample comprising:
- a different bead set specific for each of the target nucleic acid sequences, the bead set comprising (a) at least one separating bead, comprising a magnetic particle, a first codeable label comprising two or more labels, and a first target-specific probe, wherein the first codeable label is specific for the first target-specific probe, and (b) at least one detecting bead, comprising a second codeable label comprising a set of two or more labels, and a second target-specific probe, wherein the second codeable label is specific for the second target-specific probe;
- wherein the first codeable label is detectably different from the second codeable label;
- and wherein the target-specific probes in each set are suitable for ligation together when hybridized adjacent to one another on a complementary target sequence.
A method for detecting at least two different target nucleic acid sequences in a sample comprising:
- forming a ligation reaction mixture by combining the sample with a different bead set specific for each of the at least two different target nucleic acid sequences, each bead set comprising (a) at least one separating bead, comprising a magnetic particle, a codeable label comprising at least two labels, and a first target-specific probe, wherein the first codeable label is specific for the first target-specific probe, and (b) at least one detecting bead, comprising a second codeable label comprising at least two labels and a second target-specific probe, wherein the second codeable label is specific for the second target-specific probe;
- wherein the first codeable label is detectably different from the second codeable label;
- wherein the target-specific probes in each set are suitable for ligation together when hybridized adjacent to one another on a complementary target sequence;
- subjecting the ligation reaction mixture to a ligation reaction, wherein adjacently hybridizing complementary target-specific probes are ligated to one another to form a detectable complex comprising the separating bead and the detecting bead;
- and
- quantitating the at least two different target nucleic acid sequences in the sample by quantitating the detectable complex.
3 more elements
Dependencies are inferred from the claim text.