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Cancer gene methylation detection system and cancer in-vitro detection method performed in system

A methylation and cancer technology, applied in biochemical equipment and methods, combinatorial chemistry, chemical libraries, etc., can solve problems such as the impact of cancer development and development, change the tendency of cells to develop tumor phenotype, etc., to overcome tumor heterogeneity, The effect of prolonging patient survival, high sensitivity and accuracy

Pending Publication Date: 2021-04-16
BIOCHAIN BEIJING SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The results show that although DNA methylation is not dominant in every cancer, there is no doubt that changes in gene methylation modification patterns can change the developmental propensity of cells and the phenotype of tumors, thereby affecting most cancers. play an important role in the development of cancer

Method used

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  • Cancer gene methylation detection system and cancer in-vitro detection method performed in system
  • Cancer gene methylation detection system and cancer in-vitro detection method performed in system
  • Cancer gene methylation detection system and cancer in-vitro detection method performed in system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Embodiment 1

[0149] like figure 1 As shown, the implementation process of this application is as follows:

[0150] 1.1.cfDNA extraction and purification

[0151] 1.1.1. Plasma sample preparation:

[0152] Centrifuge the blood sample at 2000g for 10min at 4°C and transfer the plasma to a new centrifuge tube. Centrifuge the plasma sample at 16,000g at 4°C for 10 minutes, and proceed to the next step according to the type of collection tube used. The type of collection tube used in this experiment is other.

[0153] Table 2

[0154]

[0155] 1.1.2. Lysis and conjugation

[0156] 1.1.2.1. Prepare the binding solution / bead mixture according to the table below and mix thoroughly.

[0157] table 3

[0158]

[0159] Add an appropriate volume of plasma sample.

[0160] 1.1.2.2. Thoroughly mix the plasma sample and binding solution / bead mixture.

[0161] 1.1.2.3. Combine cfDNA fully on the rotary mixer for 10 minutes to bind the cfDNA to the magnetic beads.

[0162] 1.1.2.4. Put the b...

Embodiment 2

[0312] A case of gastric cancer sample that has been pathologically identified was detected by the Panel of the present application, and peripheral blood was collected according to the method in Example 1; the database was built and sequenced through the Illumina platform; the sequencing data was analyzed through the above-mentioned biological information to obtain the methylation level. The results are shown in Table 19 below (Table 19 shows the detected target regions greater than or equal to the methylation threshold).

[0313] Table 19

[0314] Gene CHR start termination methylation ratio target region serial number TBX15 1 119527108 119527157 0.55 Seq ID No.63 CRYGD 2 208989200 208989249 0.60 Seq ID No.64 CPE 4 166300051 166300291 0.42 Seq ID No.81 CPE 4 166300242 166300291 0.42 Seq ID No.82 PLXDC2 10 20104497 20104546 0.46 Seq ID No.97 PLXDC2 10 20104758 20104807 0.46 Seq ID No.98 P...

Embodiment 3

[0318]A sample of colorectal cancer was detected by the Panel of the present application, and peripheral blood was collected according to the method in Example 1; the library was constructed and sequenced through the Illumina platform; the sequencing data was analyzed through the above-mentioned biological information analysis process to obtain the methylation level, and the results are as follows Shown in Table 20 (Table 20 shows the detected target regions greater than or equal to the methylation threshold).

[0319] Table 20

[0320] Gene CHR start termination methylation ratio target region serial number TBX15 1 119527108 119527157 0.55 Seq ID No.63 CRYGD 2 208989200 208989249 0.60 Seq ID No.64 C6orf155 6 72130359 72130408 0.56 Seq ID No.87 C6orf155 6 72130553 72130602 0.71 Seq ID No.88 C6orf155 6 72130641 72130690 0.65 Seq ID No.89 C6orf155 6 72130755 72130804 0.69 Seq ID No.90 SHISA...

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Abstract

Disclosed herein is a system for detecting cancer methylation. The system comprises a sample collection module for collecting a subject sample, a DNA extraction module used for extracting and purifying DNA in the sample, a library building module used for building a DNA library for sequencing for the purified DNA sample, a conversion module used for converting the constructed DNA library by using a bisulfite, a pre-PCR amplification module used for carrying out pre-PCR amplification on the DNA library subjected to bisulfite conversion, a hybrid capture module used for carrying out hybrid capture on the sample subjected to the pre-PCR amplification by utilizing a probe composition, a post-PCR amplification module used for amplifying a product captured by hybridization by utilizing PCR, a sequencing module used for carrying out high-throughput next-generation sequencing on a product subjected to hybridization capture after PCR amplification, a data analysis module used for analyzing the sequencing data and determining the methylation level of the sample, and an interpretation module used for interpreting the illness condition of the patient based on the methylation level of the sample.

Description

technical field [0001] This application relates to a cancer gene methylation detection system, in particular to a system based on high-throughput sequencing (NGS) to detect changes in the methylation level of free DNA in three types of luminal organ tumors, including esophageal cancer, gastric cancer, and colorectal cancer. As well as an in vitro detection method for cancer performed in the system. Background technique [0002] High-throughput sequencing (NGS) technology is a revolutionary innovation in the field of modern genomics research. This technology can simultaneously perform sequence analysis on tens to millions of DNA molecules, which marks the arrival of the post-genome era. Through the control of the sequencing depth, different goals such as de novo sequencing and resequencing can be achieved, and the sequences of the genome, transcriptome, and methylome can also be analyzed through different pre-processing. [0003] The current clinical gene testing techniques ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6869
CPCC12Q1/6886C12Q1/6869C40B40/06C40B50/06
Inventor 韩晓亮李永君吴宁宁郭媛媛王建铭
Owner BIOCHAIN BEIJING SCI & TECH
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