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Assays for single molecule detection and use thereof

  • US 10,626,450 B2
  • Filed: 08/11/2017
  • Issued: 04/21/2020
  • Est. Priority Date: 08/19/2013
  • Status: Active Grant
First Claim
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1. A method of detecting nucleic acid copy numbers in a genetic sample obtained from a subject, comprisingcontacting first and second probe sets to the genetic sample, wherein the first probe set comprises a first labeling probe and a first tagging probe, and the second probe set comprises a second labeling probe and a second tagging probe;

  • hybridizing at least parts of probes of the first and second probe sets to first and second nucleic acid regions of interest in nucleotide molecules present in the genetic sample to produce first and second probe products, respectively, wherein the first and second probe products are circular molecules;

    amplifying the first and second probe products to form first and second amplified probe products;

    labeling the first and second amplified probe products with first and second labels to produce first and second labeled amplified probe products;

    immobilizing at least parts of probes of the first and second labeled amplified probe products to a substrate at a density in which the first and second labeled probe products are optically resolvable after immobilization; and

    detecting (i) a first number of the first labeled amplified probe products, and (ii) a second number of the second labeled amplified probe products to detect the nucleic acid copy numbers.

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