Methylation modification-based tumor marker stamp-ep6

A method for detecting a tumor, wherein said method comprises:

• detecting a modification on the CpG site(s) of a polynucleotide by a tumor detection agent or kit, if the hypermethylation of a subject is detected, the subject can be identified as having a high-risk of tumor;
• wherein said polynucleotide comprises:
• (a) a polynucleotide with a nucleotide sequence as shown in SEQ ID NO:
• 1;
• (b) a fragment of the polynucleotide of (a), having at least one CpG site with modification;
• and/or (c) a nucleic acid complementary to the polynucleotide or fragment of (a) or (b).

A method of preparing a tumor detection agent, comprising:

• providing a polynucleotide, designing a detection agent for specifically detecting the modification on CpG(s) of the a target sequence which is the full length or fragment of the polynucleotide;
• wherein, said polynucleotide comprises:
• (a) a polynucleotide with a nucleotide sequence as shown in SEQ ID NO:
• 1;
• (b) a fragment of the polynucleotide of (a), having at least one CpG site with modification;
• and/or (c) a nucleic acid complementary to the polynucleotide or fragment of (a) or (b).

The method according to claim 12, wherein the primers are:

• primers shown in SEQ ID NO:
• 5 and 6;
• or
• primers shown in SEQ ID NO:
• 7 and 8.

A detection kit, comprising:

• container(s) and a detection agent in the container(s);
• wherein said detection agent specifically detects the modification of the CpG(s) of a target sequence which is the full length or fragment of the polynucleotide;
• wherein said polynucleotide comprises:
• (a) a polynucleotide with a nucleotide sequence as shown in SEQ ID NO:
• 1;
• (b) a fragment of the polynucleotide of (a), having at least one CpG site with modification;
• and/or (c) a nucleic acid complementary to the polynucleotide or fragment of (a) or (b).

A method of detecting the methylation profile of a sample in vitro, comprising:

• (i) providing the sample and extracting nucleic acid;
• (ii) detecting a modification on CPG site(s) of a target sequence in the nucleic acid of (i), wherein the target sequence is a polynucleotide comprising:
• (a) a polynucleotide with a nucleotide sequence as shown in SEQ ID NO:
• 1;
• (b) a fragment of the polynucleotide of (a), having at least one CpG site with modification;
• and/or (c) a nucleic acid complementary to the polynucleotide or fragment of (a) or (b).

An isolated polynucleotide, wherein the isolated polynucleotide comprises:

• (a) a polynucleotide with a nucleotide sequence as shown in SEQ ID NO:
• 1 or a fragment thereof, having at least one CpG site with modification;
• said modification comprises 5-methylation, 5-hydroxymethylation, 5-formylcytosine or 5-carboxylcytosine;
• and/or
• (b) a nucleic acid complementary to the polynucleotide or fragment of (a).

An isolated polynucleotide, wherein the polynucleotide is converted from an original polynucleotide, said original polynucleotide comprises:

• (a) a polynucleotide with a nucleotide sequence as shown in SEQ ID NO:
• 1;
• (b) a fragment of the polynucleotide of (a), having at least one CpG site with modification;
• and/or (c) a nucleic acid complementary to the polynucleotide or fragment of (a) or (b);
• and as compared with the sequence of the original polynucleotide, cytosine C of the CpG site(s) with modification is unchanged, and the unmodified cytosine is converted into T or U in the converted polynucleotide.