Methods for detecting epigenetic modifications
A method of detecting a predisposition to, or the incidence of, cancer in a sample comprises detecting an epigenetic change in at least one gene selected from an NDRG4/NDRG2 subfamily gene, GATA4, OSMR, GATA5, SFRP1, ADAM23, JPH3, SFRP2, APC, MGMT, TFPI2, BNIP3, FOXE1, SYNE1, SOX17, PHACTR3 and JAM3, wherein detection of the epigenetic change is indicative of a predisposition to, or the incidence of, cancer. Also described are pharmacogenetic methods for determining suitable treatment regimens for cancer and methods for treating cancer patients, based around selection of the patients according to the methods of the invention. The present invention is also concerned with improved methods of collecting, processing and analyzing samples, in particular body fluid samples. These methods may be useful in diagnosing, staging or otherwise characterizing various diseases. The invention also relates to methods for identifying, diagnosing, staging or otherwise characterizing cancers, in particular gastrointestinal cancers such as colorectal cancers, gastric cancers and oesophageal cancers. The methods of the invention relate, inter alia, to isolating and analyzing the human DNA component from faecal samples and blood-based samples.
1. A method, comprising:
(a) treating DNA obtained from a sample with a reagent that converts non-methylated cytosines to uracil;
(b) amplifying the treated DNA with primers that hybridize to CpG dinucleotides in a target sequence that is within positions 1-1000 of SEQ ID NO:524 to produce amplified DNA that comprises CpG dinucleotides; and
(c) detecting production of the amplified DNA
wherein the sample comprises one or more of a tissue sample, a fecal sample and/or a bodily fluid sample.
2. The method of claim 1 , wherein detecting the production of the amplified DNA comprises detecting hybridization of a probe that hybridizes to the amplified DNA.
3. The method of claim 2 , wherein the probe hybridizes to CpG dinucleotides in the target sequence.
4. The method of claim 2 , wherein the probe is a hydrolytic probe and hybridization of the probe to the amplified DNA is detected by detecting hydrolysis of the probe.
5. The method of claim 1 , wherein the bodily fluid sample comprises a blood sample and/or a plasma sample.