Granted Patent B2

US 12,606,872 · App. 18/096,337 · Granted Apr 21, 2026

Inventors: Pol Canal Noguer (Seville, ES); Juan Carlos Higareda Almaraz (Ljubljana, SI); Francesco Mattia Mancuso (Seville, ES); Kristi Kruusmaa (Ljubljana, SI); Alejandro Requena Bermejo (Seville, ES); Marko Chersicola (Ljubljana, SI); Pablo Pérez Martínez (Seville, ES); Pablo Antonio Camino Faillace (Seville, ES); Primož Knap (Ljubljana, SI); Vivian Erklavec Zajec (Ljubljana, SI); Špela Zavodnik (Ljubljana, SI); Carme Nolla Colomer (Seville, ES); Fernando Trincado Alonso (Seville, ES)

Assignee: Universal Diagnostics, S.A.

C12Q1/6886C12Q2600/112C12Q2600/154

Abstract

Claims (19)

1 . A method of detecting methylation statuses of markers, the method comprising:

converting unmethylated cytosines of a plurality of DNA fragments in a sample into uracils to generate a plurality of converted DNA fragments, wherein the plurality of DNA fragments were obtained from a biological sample;

sequencing the plurality of converted DNA fragments to generate a plurality of sequence reads, wherein each sequence read corresponds to a converted DNA fragment; and

detecting a methylation status for each of three or more markers identified in the sequence reads,

wherein a first of the three or more markers is a methylation locus comprising at least a portion of SEQ ID NO. 1, a second of the three or more markers is a methylation locus comprising at least a portion of SEQ ID NO. 2, and a third of the three or more markers is a methylation locus comprising at least a portion of SEQ ID NO. 3.

2 . The method of claim 1 , wherein the plurality of DNA fragments (in total) comprise at least 1 ng of DNA.

3 . The method of claim 1 , wherein the plurality of DNA fragments consist essentially of DNA fragments each of which has a length in a range from 10 bp to 800 bp.

4 . The method of claim 1 , wherein the plurality of DNA fragments consist essentially of DNA fragments each of which has a length in a range from 1000 bp to 200,000 bp.

5 . The method of claim 1 , wherein each of the plurality of sequence reads is at least 50 bp.

6 . The method of claim 1 , wherein the method further comprises detecting a methylation status for a fourth methylation locus comprising at least a portion of SEQ ID NO. 70 and a fifth methylation locus comprising at least a portion of SEQ ID NO. 72.

7 . A method of detecting methylation statuses of markers, the method comprising:

converting unmethylated cytosines of a plurality of DNA fragments in a sample into uracils to generate a plurality of converted DNA fragments, wherein the plurality of DNA fragments were obtained from a biological sample;

sequencing the plurality of converted DNA fragments to generate a plurality of sequence reads, wherein each sequence read corresponds to a converted DNA fragment; and

detecting a methylation status for each of two or more markers identified in the sequence reads,

wherein a first of the two or more markers is a methylation locus comprising at least a portion of SEQ ID NO. 70 and a second of the two or more markers is a methylation locus comprising at least a portion of SEQ ID NO. 72.

8 . The method of claim 7 , wherein the plurality of DNA fragments (in total) comprise at least 1 ng of DNA.

9 . The method of claim 7 , wherein the plurality of DNA fragments consist essentially of DNA fragments each of which has a length in a range from 10 bp to 800 bp.

10 . The method of claim 7 , wherein the plurality of DNA fragments consist essentially of DNA fragments each of which has a length in a range from 1000 bp to 200,000 bp.

11 . The method of claim 7 , wherein each of the plurality of sequence reads is at least 50 bp.

Assignments (1)

Continuity (2)

References Cited (278)