US 11,939,629 B2
Methods and systems that detect nucleic-acid targets
Andrei Gindilis, Mukilteo, WA (US)
Filed by Andrei Gindilis, Mukilteo, WA (US)
Filed on Aug. 1, 2018, as Appl. No. 16/052,538.
Claims priority of provisional application 62/539,984, filed on Aug. 1, 2017.
Prior Publication US 2019/0185920 A1, Jun. 20, 2019
Int. Cl. C12P 19/34 (2006.01); C12Q 1/6816 (2018.01); C12Q 1/6823 (2018.01); C12Q 1/6832 (2018.01); C12Q 1/686 (2018.01)
CPC C12Q 1/6832 (2013.01) [C12Q 1/6816 (2013.01); C12Q 1/6823 (2013.01); C12Q 1/686 (2013.01)] 14 Claims
 
1. A method that detects a nucleic-acid target molecule, the method comprising:
in a first stage, introducing, to recognition probes,
a sample solution containing the nucleic-acid target molecule, and
recognition restriction endonucleases that cleave a target/recognition-probe duplex to produce a first nucleic-acid product;
in a second stage, introducing, to amplification probes of two types, each bound to a solid support and each including a marker component, including a first-type amplification probe and a second-type amplification probe,
the first nucleic-acid product, and
two types of restriction endonuclease, including a second-type restriction endonuclease that cleaves a first-product/first-type-amplification-probe duplex and a third-product/first-type-amplification-probe duplex to produce a second nucleic-acid product and a third-type restriction endonuclease that cleaves a second-product/second-type-amplification-probe duplex to produce the third nucleic-acid product; and
detecting marker component no longer bound to a solid support produced by multiple cycles of product production in the second stage.