# US 9,752,197 B2 · Method for detecting methylation of colorectal cancer specific methylation marker gene for colorectal cancer diagnosis

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- Number: US 9,752,197 B2
- Document ID: US9752197B2
- Status: Granted
- Kind: B2
- Application number: 15/010,379
- Filed: 2016-01-29
- Published: 2016-05-26
- Granted: 2017-09-05
- Earliest claimed: 2010-12-16
- Assignees: Genomictree, Inc.
- Applicants: GENOMICTREE, Inc.
- Inventors: Sung Whan An; Tae Jeong Oh
- Web page: https://patentgrep.com/patent/US9752197B2

## CPC

- C12Q 1/6886: for cancer (immunoassay for cancer G01N33/575)
- C12Q 2600/112: Disease subtyping, staging or classification
- C12Q 2600/154: Methylation markers
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The present invention relates to a method for detecting methylation of the bowel-cancer-specific methylation marker GPM6A (NM_201591, glycoprotein M6A) gene in order to diagnose bowel cancer, and more specifically relates to a method for providing information for diagnosing bowel cancer by detecting the methylation of a bowel-cancer-specific marker gene that is specifically methylated in bowel cancer cells. The method for detecting methylation and a diagnostic composition, kit and nucleic-acid chip according to the present invention can be used to advantage in diagnosing bowel cancer more accurately and quickly than by normal methods as they permit bowel cancer to be diagnosed at the initial genetic transformation step and so allow early diagnosis.

## Claims

### Claim 1

A method for detecting CpG methylation of GPM6A (glycoprotein M6A) gene, the method comprising the steps of:
(a) isolating genomic DNA from a clinical sample;
(b) treating the genomic DNA or a fragment thereof with bisulfite;
(c) amplifying a methylated CpG of GPM6A gene in the bisulfite-treated genomic DNA or fragment thereof from step (b) by using primer(s) comprising sequence(s) selected from the group consisting of SEQ ID Nos: 24 to 69; and (d) determining whether the CpG GPM6A was methylated based on whether the DNA was amplified in step (c).

### Claim 2 (depends on 1)

The method of claim 1, wherein the step (c) is performed by detecting methylated CpG of the promoter or the CpG of the first intron of GPM6A gene.

### Claim 3 (depends on 2)

The method of claim 2, wherein the first intron of GPM6A gene has a sequence of SEQ ID NO: 1.

### Claim 4 (depends on 1)

The method of claim 1, wherein the detection of methylation is performed by a method selected from the group consisting of PCR, methylation-specific PCR, real-time methylation-specific PCR, quantitative PCR, pyrosequencing, and bisulfite sequencing.

### Claim 5 (depends on 1)

The method of claim 1, wherein the clinical sample is selected from the group consisting of a tissue, cell, blood, blood plasma, serum, feces, and urine from a patient suspected of cancer or a subject to be diagnosed.

### Claim 6 (depends on 1)

The method of claim 1, further comprising using probe(s) capable of hybridizing with a methylated CpG of GPM6A comprising at least one or more CpG dinucleotide in a region which hybridizes to the methylated CpG of GPM6A.

### Claim 7 (depends on 6)

The method of claim 6, wherein the probe(s) comprises sequence(s) selected from the group consisting of SEQ ID NOS: 70-92.