IP Library › Patent Application 18544344
Patent Application
App. No. 18/544,344

Ultra-Sensitive Platform for Nucleic acid detection using a novel method, Scanning Digital polymerase chain reaction (PCR)

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Quick Facts
Patent No.
US None
App. No.
18/544,344
Abstract

A method for analyzing a target nucleic acid includes diluting nucleic acid targets and filling pico to femto-liter sized wells such that they contain a single target nucleic acid and one or more amplification reagents, amplifying the target in the individual wells, distinguishing wells containing amplicon from the target and amplicon from a variant of the target generated by polymerase error by using two differently labeled-hybridization probes, one hybridizing to the target and one hybridizing to a specific variant of the target; and analyzing target amplicons.

Claims (18)

1 . A method for analyzing a target nucleic acid, the method comprising the steps of:

diluting nucleic acid targets and filling pico to femto-liter sized wells such that

they

contain a single target nucleic acid and one or more amplification reagents;

amplifying the target in the individual wells;

distinguishing wells containing amplicon from the target and amplicon from a variant of the target generated by polymerase error by using two differently labeled-hybridization probes, one hybridizing to the target and one hybridizing to a specific variant of the target; and

analyzing target amplicons.

2 . The method according to claim 1 , wherein said amplifying step is a polymerase chain reaction and the one or more amplification reagents includes one or more primer pairs.

3 . The method according to claim 1 , wherein said distinguishing step comprises scanning a laser across each well individually.

4 . The method according to claim 1 , wherein said analyzing step comprises detecting said amplicons by hybridization to detectably-labeled probes.

5 . The method according to claim 1 , wherein said analyzing step is conducted on amplicon from wells that were not distinguished in said distinguishing step.

6 . The method according to claim 1 , wherein said analyzing step comprises:

determining a number of wells that contain only wild-type target;

determining a number of wells that contain only a variant of the target.

7 . The method according to claim 6 , wherein presence of wells containing only said variant is indicative of a disease.

8 . The method according to claim 7 , wherein the disease is cancer.

9 . The method according to claim 6 , wherein the variant is an allelic variant.

10 . The method according to claim 9 , wherein the allelic variant is a single nucleotide polymorphism.