Detection of a panel of urine DNA markers for HCC screening and disease management

摘要

Provided herein is a method for detecting the presence or absence of a cancer in a biological sample of an individual, by determining the level of mutation and methylation of one or more genes from a group of genes comprising TP53, CTNNB1, hTERT, RASSF1A, GSTP1, p16, p15 and SFRP-1. Also provided herein is an assay to detect p53 mutations suitable for DNA isolated from biological body fluid in order to screen cancer patients.;Also provided is a method for detecting the presence or absence of a liver cancer in an individual by determining the level of methylation.;Also provided is a suitable method for detecting the presence or absence of a liver cancer in an individual by determining the level of methylation of the promoter of the GSTP1 gene in body fluid such as urine or blood.;Also provided is a suitable method for detecting the presence or absence of a liver cancer in an individual by determining the level of methylation of the promoter of the RASSF1A gene in body fluid such as urine or blood.

主权项

1. A method of detecting the presence or absence of hepatocellular carcinoma (HCC) in a subject in need thereof, comprising:
(1) preparing a DNA sample from a urine sample of the subject; (2) determining a level of methylation of RASSF1A from the DNA sample of the subject by a two-step short-amplicon methylation specific PCR (shMSP), wherein the two-step short-amplicon methylation specific PCR (shMSP) uses primers of the nucleotide sequences as set forth in SEQ ID NO: 15 and SEQ ID NO:16 for a first step PCR and SEQ ID NO:17 and SEQ ID NO:18 for a second step PCR; (3) comparing the level of methylation of RASSF1A with a level of methylation of RASSF1A in one or more control samples from subjects known not to have HCC; and (4) detecting the presence or absence of HCC, with elevated methylation levels in RASSF1A of the individual as compared to the level of methylation in RASSF1A in the one or more control samples indicating the presence of HCC, and the absence of elevated methylation levels indicating the absence of HCC.