	US 12,188,093 B2
	Detecting cholangiocarcinoma
John B. Kisiel, Rochester, MN (US); David A. Ahlquist, Rochester, MN (US); William R. Taylor, Lake City, MN (US); Douglas W. Mahoney, Elgin, MN (US); and Tracy C. Yab, Rochester, MN (US)
Assigned to Mayo Foundation for Medical Education and Research, Rochester, MN (US)
Filed by Mayo Foundation for Medical Education and Research, Rochester, MN (US)
Filed on Dec. 14, 2020, as Appl. No. 17/120,518.
Application 17/120,518 is a continuation of application No. 16/242,695, filed on Jan. 8, 2019, granted, now 10,900,090.
Application 16/242,695 is a continuation of application No. 14/864,158, filed on Sep. 24, 2015, granted, now 10,184,154, issued on Jan. 22, 2019.
Claims priority of provisional application 62/055,737, filed on Sep. 26, 2014.
Prior Publication US 2021/0102264 A1, Apr. 8, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6886 (2018.01)

CPC C12Q 1/6886 (2013.01) [C12Q 2600/112 (2013.01); C12Q 2600/154 (2013.01)]

2 Claims

1. A method for measuring the methylation level of one or more CpG sites, the method comprising:

a) extracting genomic DNA from a biological sample of a human individual suspected of having or having a neoplasm, wherein the neoplasm is intra-hepatic cholangiocarcinoma or extra-hepatic cholangiocarcinoma;

b) treating the extracted genomic DNA with bisulfite;

c) amplifying the bisulfite-treated genomic DNA with primers specific for EMX1, and at least one of ZNF781, HOXA1, and CYP26C1; and

d) measuring the methylation level of the one or more CpG sites in the EMX1 and the at least one of ZNF781, HOXA1, and CYP26C1 by methylation-specific PCR, quantitative methylation-specific PCR, methylation sensitive DNA restriction enzyme analysis or bisulfite genomic sequencing PCR.