Polymerization-induced separation assay using recognition pairs

A method employing recognition pairs of reactants for determining the presence or amount of a nucleic acid analyte in a sample suspected of containing said analyte comprising:

• (a) labeling as a first recognition reactant a first single stranded polynucleotide probe capable of specifically binding to the analyte with either of an addition polymerizable organic monomer or reporter;
• (b) contacting the sample with the labeled first probe to form a first probe-analyte complex;
• (c) labeling as a second recognition reactant a second single stranded polynucleotide probe capable of specifically binding to the analyte with the alternative label not used for the first probe;
• (d) contacting the sample sequentially or simultaneously with the labeled second probe to form a first probe-analyte-second probe ternary complex;
• (e) separating the resultant ternary complex from free reporter-labeled probe by initiating polymerization of the monomer-labeled probe in the presence of an unlabeled addition polymerizable organic monomer;
• and
• (f) detecting the incorporation of reporter-labeled probe into said polymerized ternary complex to determine the presence of the analyte.

A method for detecting the presence or amount of association between two analytes in a sample suspected of containing both analytes comprising:

• (a) labeling a first recognition reactant capable of specifically binding to a first analyte with either of addition polymerizable monomer or reporter;
• (b) contacting said sample with the labeled first recognition reactant to form a first recognition reactant-first analyte complex;
• (c) labeling a second recognition reactant capable of specifically binding to the second analyte with the alternative label not used for the first recognition reactant;
• (d) contacting said sample sequentially or simultaneously with the labeled second reactant to form a second recognition reactant-second analyte complex;
• (e) initiating polymerization of the monomer-labeled recognition reactant whereby the first complex is separated from the second complex unless they are associated;
• and
• (f) detecting the incorporation of reporter-labeled recognition reactant into said polymerized complex to determine the presence or amount of analyte association.

A method for determining the presence or amount of an analyte in a sample suspected of containing said analyte comprising:

• (a) labeling analyte with either of an addition polymerizable organic monomer or reporter;
• (b) contacting the sample with the labeled analyte to form a mixture;
• (c) labeling a recognition reactant capable of specifically binding to the analyte or the labeled analyte with the alternative label not used in step (a);
• (d) contacting the mixture with the labeled recognition reactant to form recognition reactant-analyte complex;
• (e) separating monomer-containing recognition reactant-analyte complex from recognition reactant-analyte complex not labeled with monomer by initiating polymerization of the monomer-labeled reactant in the presence of an unlabeled addition polymerizable organic monomer;
• and
• (f) detecting the incorporation of reporter into said polymerized complex and relating this incorporation to the analyte present in the sample.

A method employing recognition pairs of reactants for determining the presence or amount of a nucleic acid analyte in a sample suspected of containing said analyte, comprising:

• (a) labeling a first antibody reactant capable of specifically binding to the nucleic acid analyte with either of an addition polymerizable organic monomer or reporter;
• (b) contacting the sample with the labeled reactant to form an antibody reactant-analyte complex;
• (c) labeling a nucleic acid probe capable of specifically binding to the nucleic acid analyte with the alternative label not used for the antibody reactant;
• (d) contacting the sample either sequentially or simultaneously with the nucleic acid probe to form an antibody reactant-analyte-nucleic acid probe ternary complex;
• (e) separating the resultant ternary complex from free reporter-labeled recognition reactant by initiating polymerization of the monomer-labeled recognition reaction;
• and
• (f) detecting the incorporation of reporter-labeled recognition reactant into said polymerized ternary complex and relating this incorporation to the analyte present in the sample.

A method employing recognition pairs of nucleic acids for determining the presence or amount of a nucleic acid analyte in a sample suspected of containing said analyte, comprising:

• (a) labeling as a first recognition reactant a first single stranded polynucleotide probe capable of specifically binding to the analyte with either of an addition polymerizable organic monomer or reporter;
• (b) contacting the sample with the labeled reactant to form a first probe-analyte complex;
• (c) labeling as a second recognition reactant a second single stranded polynucleotide probe capable of specifically binding to the analyte with the alternative label not used for the first probe;
• (d) contacting the sample either sequentially or simultaneously with the labeled second probe to form a first probe-analyte-second probe ternary complex;
• (e) separating the resultant ternary complex from free reporter-labeled probe by initiating polymerization of the monomer-labeled probe;
• and
• (f) detecting the incorporation of reporter-labeled probe into said polymerized ternary complex and relating this incorpration to the analyte present in the sample.