Colorectal cancer polygene methylation joint detection test kit and application thereof

A detection kit and technology for colorectal cancer, applied in the field of molecular biology, can solve problems such as reducing the positive detection rate, achieve high sensitivity, strong specificity, and improve screening efficiency

Inactive Publication Date: 2020-08-14
山东龙奥生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] At present, there are many gene methylation status detection kits for colorectal cancer detection, but most of the kits only target the methylation detection of a single gene such as the SEPT9 gene, or the detection of a small number of 2 or 3 genes. This greatly reduces the positive detection rate

Method used

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  • Colorectal cancer polygene methylation joint detection test kit and application thereof
  • Colorectal cancer polygene methylation joint detection test kit and application thereof
  • Colorectal cancer polygene methylation joint detection test kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] 1 Materials and methods

[0056] 1.1 sample

[0057] Stool and blood samples were collected from 52 patients with colorectal cancer diagnosed by colonoscopy, and from 46 normal healthy people. There was no significant difference in ethnicity and regional origin between the disease group and the control group, all of them were Han nationality.

[0058] 1.2 Genomic DNA extraction

[0059] 1.2.1 Sample pretreatment

[0060] Take 200mg-300mg of stool sample, add 1mL of impurity-removing solution, shake vigorously for 1min, centrifuge at 12000rpm for 10min, and take 500μL of supernatant into a 2mL centrifuge tube.

[0061] The blood sample was centrifuged at 3000 rpm for 10 minutes, and 300 μL of the supernatant was taken into a 2 mL centrifuge tube.

[0062] 1.2.2 Sample Lysis

[0063] Add 10 μL of proteinase K and 300 μL of lysate to the above 2 mL centrifuge tube, vortex to mix, and incubate at 60°C for 10 min.

[0064] 1.2.3 DNA adsorption binding

[0065] Add 500...

Embodiment 2

[0115] 1 Materials and methods

[0116] 1.1 Sample Collection

[0117] Stool samples from 52 patients with colorectal cancer confirmed by colonoscopy and 46 normal healthy people were collected. There was no significant difference in ethnicity and regional origin between the disease group and the control group, all of them were Han nationality.

[0118] 1.2 Genomic DNA extraction

[0119] Take 200mg-300mg of stool sample, add 1mL of impurity-removing solution, shake vigorously for 1min, centrifuge at 12000rpm for 10min, and take 500μL of supernatant into a 2mL centrifuge tube.

[0120] All the other steps are the same as 1.2 steps in Example 1.

[0121] 1.3 Bisulfite conversion

[0122] The steps are the same as the 1.3 steps in Example 1.

[0123] 1.4 Methylation Detection

[0124] 1.4.1 Reagent preparation

[0125] Colorectal cancer multi-gene methylation joint detection kit, including PCR reaction solution A (including primers and probes of SEPT9 gene, SDC2 gene and ...

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Abstract

The invention discloses a colorectal cancer polygene methylation joint detection test kit and application thereof, and belongs to the technical field of molecular biology. The test kit provided by theinvention provides primers and probes for CpG island methylation detection of SEPT9, SDC2, SFRP2, NDRG4, SPG20 and FBN1 genes, and gene methylation detection is carried out by adopting a bisulfite fluorescent quantitative PCR method. The test kit provides an effective means and basis for evaluating the potential possibility of colorectal cancer, and has a wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to a colorectal cancer SEPT9, SDC2, SFRP2, NDRG4, SPG20, FBN1 multi-gene methylation combined detection kit, detection method and application thereof. Background technique [0002] DNA methylation is an epigenetic modification of genes. DNA methylation can turn off the activity of some genes. DNA methylation occurs on the cytosine of CpG, and under the action of DNA methyltransferases (DNMTs), the cytosine at the 5' end of the CpG dinucleotide is converted into 5' methylcytosine. This DNA modification method regulates the expression of genes without changing the DNA sequence, and plays an important role in maintaining the normal function of cells, embryonic development and human tumorigenesis. [0003] Colorectal cancer (CRC) is the third most common cancer in the world. In 2012, 1.36 million cases of CRC were diagnosed worldwide. This figure is still growing a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/6886C12Q1/686C12Q2600/154C12Q2600/166C12Q2600/16
Inventor 汪媛媛黄任平李本涛李仕亮
Owner 山东龙奥生物技术有限公司
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