	US 11,667,973 B2
	Endosialin (CD248) as epigenetic marker for the identification of immune cells, in particular naïve CD8+ t-cells
Sven Olek, Berlin (DE); and Janika Josephin Schulze, Berlin (DE)
Assigned to Precision for Medicine GmbH, Berlin (DE)
Appl. No. 16/758,896
Filed by Precision for Medicine GmbH, Berlin (DE)
PCT Filed Oct. 25, 2018, PCT No. PCT/EP2018/079290 § 371(c)(1), (2) Date Apr. 24, 2020, PCT Pub. No. WO2019/081642, PCT Pub. Date May 2, 2019.
Claims priority of application No. 102017125150.2 (DE), filed on Oct. 26, 2017.
Prior Publication US 2020/0370117 A1, Nov. 26, 2020
Int. Cl. C12Q 1/6881 (2018.01)

CPC C12Q 1/6881 (2013.01) [C12Q 2600/154 (2013.01)]

15 Claims

1. A method for producing an amplicon from a human endosialin (CD248) gene, the method comprising:

a) bisulfite treating isolated genomic DNA from a human immune cell sample to generate bisulfite treated DNA, and

b) amplifying a region of the CD248 gene from the bisulfite treated DNA to thereby produce the amplicon, wherein the amplifying is performed with a polymerase chain reaction (PCR) using methylation-specific primers that amplify a region of SEQ ID NO: 2 and the amplicon comprises thymine-guanine (TG) at nucleotide positions 274 and 275, 292 and 293, and 296 and 297 of SEQ ID NO: 2.