The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Oct. 21, 2025

Filed:

Oct. 14, 2019
Applicant:

The Johns Hopkins University, Baltimore, MD (US);

Inventors:

Saraswati Sukumar, Columbia, MD (US);

Mary Jo Fackler, Hunt Valley, MD (US);

Wei Wen Teo, Johor Bahru Johor, MY;

Zoila Areli Lopez Bujanda, Bethesda, MD (US);

Antonio Wolff, Baltimore, MD (US);

Assignee:

The Johns Hopkins University, Baltimore, MD (US);

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12Q 1/6883 (2018.01); C12P 19/34 (2006.01); C12Q 1/686 (2018.01); C12Q 1/6886 (2018.01);
U.S. Cl.
CPC ...
C12Q 1/6883 (2013.01); C12P 19/34 (2013.01); C12Q 1/686 (2013.01); C12Q 1/6886 (2013.01); C12Q 2523/125 (2013.01); C12Q 2525/161 (2013.01); C12Q 2525/185 (2013.01); C12Q 2525/204 (2013.01); C12Q 2600/118 (2013.01); C12Q 2600/154 (2013.01); C12Q 2600/16 (2013.01);
Abstract

A cMethDNA method quantitatively detects tumor DNA (or other circulating DNAs) in fluids such as serum or plasma at the lowest copy number. Unique compared to any other PCR-based assay, a small number of copies of a synthetic polynucleotide standard (STDgene) is added to an aliquot of patient serum. A cocktail of standards for a plurality of genes of interest (TARGETgene) is added to a sample of serum. Once total DNA is purified and processed, a PCR (multiplex step) is performed wherein the STDgene and the TARGETgene are co-amplified with the same external primer set. In the second nested PCR step, amplicons present in a dilution of the first PCR reaction are subjected to real time PCR, and quantified for each gene in one well by two-color real-time PCR. Products are calculated by absolute quantitation with internal primer sets specific for the methylated TARGETgene and associated STDgene.


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