# US 7,608,402 B2 · DNA methylation specific signal amplification

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- Number: US 7,608,402 B2
- Document ID: US7608402B2
- Status: Granted
- Kind: B2
- Application number: 12/008,292
- Filed: 2008-01-10
- Published: 2009-07-16
- Granted: 2009-10-27
- Earliest claimed: 2008-01-10
- Inventors: Weiwei Li; Jessica Li
- Web page: https://patentgrep.com/patent/US7608402B2

## CPC

- C12Q 1/6853: using modified primers or templates
- 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

This invention is related to a method for rapidly detecting gene specific methylation through signal amplification using the epi-barcode after hybrid capture of the methylated DNA sequence.

## Claims

### Claim 1

A method for amplifying methylation specific signal generated from methylated DNA sequence in a DNA sample comprising steps of: a) binding of a nanobead adaptor consisted of a carrier bead, a bridge antibody and holder oligonucleotides to anti-5-methylcytosine antibody bound to said methylated DNA sequence; b) binding of an epi-barcode consisting of polynucleotides therefore containing 5-methylcytosine to nanobead adaptor; and c) detection of the presence of the epi-barcode with the anti-5-methylcytosine antibody wherein the amount of the epi-barcode bound to nanobead adaptor determines the amplification intensity of methylation specific signal generated from said methylated DNA sequence in the DNA sample.

### Claim 2 (depends on 1)

The method according to claim 1 wherein said carrier bead is a polypropylene bead or a magnetic bead in the size of 5 nm to 900 nm in diameter and coated with streptavidin, avidin or neutravidin.

### Claim 3 (depends on 1)

The method according to claim 1 wherein said bridge antibody is anti-mouse, or anti-rabbit, or anti-goat, or anti-sheep, or anti-chicken IgG or IgM and labeled with biotin.

### Claim 4 (depends on 1)

The method according to claim 1 wherein said holder oligonucleotides are oligo dT, or oligo dG, or oligo dC, or oligo dA with a length of 10 to 100 nucleotides and labeled with biotin.

### Claim 5 (depends on 1)

The method according to claim 1 wherein said nanobead adaptor is formed through immobilizing bridge antibody and holder oligonucleotides to the carrier bead.

### Claim 6 (depends on 1)

The method according to claim 1 wherein said epi-barcode is the synthesized poly (dI-dC)•(dI-dC), or poly (dC-dG)•((dC-dG) with a length of 3-10,000 bases or base pairs containing 1-10,000 5-methylcytosines.

### Claim 7 (depends on 1)

The method according to claim 1 wherein said epi-barcode is labeled with biotinylated dATP, dCTP, dGTP, dTTP, or dUTP or tailed with poly dA, or poly dT, or poly dC, or poly dG.