FRET-BASED ANALYTES DETECTION AND RELATED METHODS AND SYSTEMS

摘要

FRET-based analytes detection and related methods and systems are described where a pair of FRET labeled primers and/or oligonucleotides are used that are specific for target sequences located at a distance up to four time the Förster distance of the FRET chromophores presented on the FRET labeled primers and/or oligonucleotides one with respect to the other in one or more polynucleotide analyte; in particular the pair of FRET labeled primers and/or oligonucleotides is combined with a sample and subjected to one or more polynucleotide amplification reactions before measuring FRET signals from at least one FRET chromophore.

主权项

1. A kit to detect at least one polynucleotide analyte in a sample, the kit comprising
at least one primer pair formed by a forward primer attaching a first FRET chromophore and a reverse primer attaching a second FRET chromophore, wherein the first FRET chromophore and the second FRET chromophore are selected to provide an energy transfer from one to another when located at a Förster distance one with respect to the another thus forming a FRET donor-acceptor chromophore pair; the forward primer has a sequence specific for a first single strand target polynucleotide within the at least one polynucleotide analyte the reverse primer has a sequence specific for a second single strand target polynucleotide within the at least one polynucleotide analyte the first single strand target polynucleotide and the second single strand target polynucleotide are located within the at least only polynucleotide analyte so that upon specific binding of the forward primer with the first target polynucleotide and specific binding of the reverse primer with the second target polynucleotide, the first FRET chromophore and the second FRET chromophore are located at a distance up to four time the Förster distance one with respect to the other.