Compositions and methods for quantifying a nucleic acid sequence in a sample

A non-amplifiable detectable polynucleotide probe, the probe comprising from 5′ to 3′:

• a fluorescent reporter, a forward template target recognition region, a polymerase-arresting molecule incorporated into the polynucleotide, a reverse template target recognition region, and a quencher molecule, wherein the fluorescent reporter and quencher molecule are separated by more than 10 bases, wherein the probe comprises one or more modified nucleotides that have enhanced binding affinity to a complementary nucleotide in a target sequence, and wherein the polymerase-arresting molecule is selected from the group consisting of a polynucleotide adduct, 2′OMe RNA amidite, and thymidine glycol.

A non-amplifiable detectable polynucleotide probe, the probe comprising from 5′ to 3′:

• a fluorescent reporter, a forward template target recognition region, a polymerase-arresting molecule incorporated into the polynucleotide, a reverse template target recognition region, and a quencher molecule, wherein the fluorescent reporter and quencher molecule are separated by more than 10 bases, wherein the polymerase-arresting molecule comprises a C-3 spacer, wherein the probe comprises one or more locked nucleic acids (LNAs), 2′ Fluoro amidites, or 2′OMe RNA amidites, and wherein the polymerase-arresting molecule is selected from the group consisting of a polynucleotide adduct, 2′OMe RNA amidite, and thymidine glycol.