	US 12,247,258 B2
	Epigenetic method to detect and distinguish IPEX and IPEX-like syndromes, in particular in newborns
Sven Olek, Berlin (DE); and Rosa Bacchetta, Menlo Park, CA (US)
Assigned to Precision for Medicine GmbH, Berlin (DE); and The Board of Truees of the Leland Stanford Junior University, Stanford, CA (US)
Appl. No. 17/255,657
Filed by Precision for Medicine GmbH, Berlin (DE); and The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
PCT Filed Jul. 3, 2019, PCT No. PCT/EP2019/067918 § 371(c)(1), (2) Date Dec. 23, 2020, PCT Pub. No. WO2020/007951, PCT Pub. Date Jan. 9, 2020.
Claims priority of provisional application 62/694,149, filed on Jul. 5, 2018.
Prior Publication US 2023/0183804 A1, Jun. 15, 2023
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/686 (2018.01); C12Q 1/6883 (2018.01)

CPC C12Q 1/6883 (2013.01) [C12Q 1/686 (2013.01); C12Q 2600/112 (2013.01); C12Q 2600/154 (2013.01)]

16 Claims

1. A method for producing and detecting a panel of amplicons from a human subject having immunodysregulation polyendocrinopathy enteropathy X-linked (IPEX) or IPEX-like disease, the method comprising:

a) bisulfite treating isolated genomic DNA from a cell sample of a human subject to convert unmethylated cytosines into uracils;

b) producing the panel of amplicons by:

i) amplifying from the bisulfite treated DNA a human gene region for FOXP3, a human CD4 gene region, a human CD3 gene region, and a housekeeping gene region;

and

ii) amplifying a nucleic acid template comprising unconverted glyceraldehyde 3-phosphate dehydrogenase (GAPDH) artificial region with inverted CpG dinucleotides (GAP[GC]), and

c) detecting the panel of amplicons.