Polynucleotide assay reagent and method

Claim 1

A diagnostic system for determination of a single-stranded polynucleotide analyte containing a selected heteromeric target sequence of bases, said system comprising: a diagnostic reagent composed of a solid support, and attached to the solid support, multiple polymer molecules, each composed of a heteromeric sequence of base-complementary recognition moieties selected from the group consisting of purine and pyrimidine heterocycles adapted to hydrogen-bond to corresponding, contiguous bases in the target sequence, under selected binding conditions, and an unbranched, substantially uncharged backbone, composed of subunit backbone moieties, supporting the recognition moieties at positions which allow hydrogen bonding between the recognition moieties and the corresponding bases in the target sequence, where the subunit backbone moieties are chosen from a group consisting of cyclic moieties, of 5 to 7 atoms in length, and acyclic moieties, of 4 to 6 atoms in length; and, molecules of a reporter, where said reporter is composed of an oligocationic tail adapted to bind electrostatically to the charged backbone of the polynucleotide analyte, and attached to the tail, one or more reporter groups adapted to produce a signal by which the presence of the reporter can be detected.

Claim 10

A method for determination of a polynucleotide analyte containing a selected target base sequence, said method comprising providing a diagnostic reagent composed of a solid support and, attached to the support, multiple polymer molecules, each composed of a heteromeric sequence of base-complementary recognition moieties selected from the group consisting of purine and pyrimidine heterocycles effective to hydrogen-bond to corresponding, contiguous bases in the target sequence, under selected binding conditions, and an unbranched, substantially uncharged, substantially stereoregular backbone, composed of subunit backbone moieties, supporting the recognition moieties at positions which allow hydrogen bonding between the moieties and the corresponding bases in the target sequence, where the subunit backbone moieties are chosen from the group consisting of cyclic moieties, of 5 to 7 atoms in length, and acyclic moieties, of 4 to 6 atoms in length; and, mixing the reagent and an analyte-containing sample under such selected conditions, to produce sequence-specific binding of the analyte to the reagent polymers, providing a reporter composed of an oligocationic tail adapted to bind electrostatically to the charged backbone of the polynucleotide analyte, and attached to the tail, one or more reporter groups adapted to produce a signal by which the presence of the reporter can be detected, binding the reporter to analyte which has bound by sequence-specific binding to the reagent, and determining the extent of reporter binding to the analyte.

Claim 11

A method for determination of a polynucleotide analyte containing a selected target base sequence, said method comprising providing a diagnostic reagent composed of a solid support and, attached to the support, multiple polymer molecules, each composed of a heteromeric sequence of base-complementary recognition moieties selected from the group consisting of purine and pyrimidine heterocycles effective to hydrogen-bond to corresponding, contiguous bases in the target sequence, under selected binding conditions, and an unbranched, substantially uncharged, substantially stereoregular backbone, composed of subunit backbone moieties, supporting the recognition moieties at positions which allow hydrogen bonding between the moieties and the corresponding bases in the target sequence, where the subunit backbone moieties are chosen from the group consisting of cyclic moieties, of 5 to 7 atoms in length, and acyclic moieties, of 4 to 6 atoms in length; and, mixing the reagent and an analyte-containing sample under such selected conditions, to produce sequence-specific binding of the analyte to the reagent polymers, providing a reporter composed of an oligocationic tail adapted to bind electrostatically to the charged backbone of the polynucleotide analyte, and attached to the tail, one or more reporter groups adapted to produce a signal by which the presence of the reporter can be detected, binding the reporter to analyte which has bound by sequence-specific binding to the reagent, and determining the extent of reporter binding to the analyte.