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US 9,957,575 B2Granted
Methods for detecting CpG methylation and for diagnosing cancer
AssigneeEpigenomics AG
InventorsDenise Kottwitz, Jörn Lewin, Anne Schlegel, Reimo Tetzner
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Abstract
The present invention relates to the field of pharmacogenomics and in particular to detecting the presence or absence of hypermethylated DNA. The detection of CpG methylation in marker DNA is useful for the diagnosis of cancers and the invention provides improved methods for this purpose. These improved methods allow in particular for a more sensitive detection of methylated marker DNA with high backgrounds of unmethylated marker DNA.
Claims14
A method for detecting hypermethylation of a target DNA in a sample comprising genomic DNA, the method comprising the steps of:
- (a) obtaining a sample comprising genomic DNA;
- (b) converting unmethylated cytosine to another base in the genomic DNA using a converting agent, wherein the genomic DNA is at least partially fragmented;
- (c) amplifying the converted genomic DNA using a pair of methylation non-specific primers comprising the nucleic acid sequences of SEQ ID NOs:
- 121 and 122 and at least one methylation specific blocker, wherein the methylation specific blocker comprises the nucleic acid sequence of SEQ ID NO:
- 123 or 124, and wherein the hypermethylated target DNA is a region of genomic human SHOX2 DNA;
- and
- (d) detecting the amplicon, wherein detecting the amplicon detects hypermethylation of the target DNA in the sample.
4 more elements
Dependencies are inferred from the claim text.