US 10,889,863 B2
FRET-based analytes detection and related methods and systems
Emil P. Kartalov, Pasadena, CA (US); Aditya Rajagopal, Irvine, CA (US); Axel Scherer, Barnard, VT (US); and Mark D. Goldberg, Alta Loma, CA (US)
Assigned to CALIFORNIA INSTITUTE OF TECHNOLOGY, Pasadena, CA (US)
Filed by CALIFORNIA INSTITUTE OF TECHNOLOGY, Pasadena, CA (US)
Filed on Aug. 13, 2018, as Appl. No. 16/102,583.
Application 16/102,583 is a continuation of application No. 14/633,094, filed on Feb. 26, 2015, granted, now 10,077,475.
Application 14/633,094 is a continuation in part of application No. 14/162,725, filed on Jan. 23, 2014, abandoned.
Claims priority of provisional application 61/944,856, filed on Feb. 26, 2014.
Claims priority of provisional application 61/756,343, filed on Jan. 24, 2013.
Prior Publication US 2019/0127799 A1, May 2, 2019
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/70 (2006.01); C12Q 1/6883 (2018.01); C12Q 1/6851 (2018.01)
CPC C12Q 1/6883 (2013.01) [C12Q 1/68 (2013.01); C12Q 1/6851 (2013.01); C12Q 1/701 (2013.01); C12Q 1/703 (2013.01); C12Q 2600/156 (2013.01); Y10T 436/143333 (2015.01)] 38 Claims
 
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 Forster 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 one 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 times the Forster distance one with respect to the other; and
wherein at least one or both the first FRET chromophore and the second FRET chromophore is attached within 10 bases of a 5′ end of at least one or both the forward primer and the reverse primer and the forward primer is not complementary to the reverse primer.