US 2018/0002743 A1 · FLUOROGENIC PROBES AND THEIR USE IN QUANTITATIVE DETECTION OF TARGET RNA SEQUENCES

Abstract

Disclosed are compositions, reagents, methods, assays, and kits for quantitative and sensitive detection of target ribonucleic acid (RNA) sequences, particularly target RNA sequences that contain RNA spliced sites, hairpin stem-loops or other topological configurations of RNA secondary, tertiary, and quaternary structure, as well as linear RNA sequences. In some embodiments, fluorogenic modified-backbone oligonucleotide probes are employed for specific sequence hybridization across the target site, followed by chemical autoligation to produce a fluorescent molecule within the ligated probes. The autoligation detection reaction is performed isothermally or through thermocycling.

Claims20

Dependent claims

A fluorogenic nucleic acid composition for quantitative detection of a target RNA sequence in a test sample comprising at least one pair of oligonucleotide probes with an upstream first probe and a downstream second probe complementary to and capable of specifically binding the target RNA in an upstream and downstream portion of the target RNA, respectively,

wherein the at least one pair of oligonucleotide probes bind to a target RNA,
wherein both probes are covalently bound to a nonfluorescent moiety,
wherein the nonfluorescent moiety bound to the first probe chemically reacts with the nonfluorescent moiety on the second probe to form a fluorescent moiety covalently connecting the two probes when both probes of the probe set hybridize to the target RNA sequence;
and
wherein quantitative detection of target RNA may be performed by detecting the fluorescent signal generated.

2 more elements

Dependencies are inferred from the claim text.

Fluorogenic probes and their use in quantitative detection of target rna sequences

Life of this patent

Fluorogenic probes and their use in quantitative detection of target rna sequences

Life of this patent

Abstract

Claims20