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First Claim
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1. 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.
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Abstract
The invention relates to methods and compositions for the detection of targets in a sample.
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Citations
38 Claims
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1. A method for quantitating a target comprising;
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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. - View Dependent Claims (2, 15, 25, 26)
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3. A method for quantitating at least two different particular targets comprising;
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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. - View Dependent Claims (4)
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5. A method for quantitating at least two different target nucleic acid sequences in a sample comprising:
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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 for each different target nucleic acid sequence. - View Dependent Claims (6, 7, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21, 22, 23, 24, 32, 33)
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8. A method for detecting at least two different target nucleic acid sequences in a sample comprising:
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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. - View Dependent Claims (27, 28, 29, 30, 31)
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34. 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.- View Dependent Claims (35, 36, 37, 38)
Specification