Probe composition for detecting three lumen organ tumors

A composition and probe technology, applied in the field of DNA sequence probe composition, can solve problems such as the influence of cancer occurrence and development, change the tumor phenotype of cell development tendency, and achieve the goal of overcoming tumor heterogeneity and saving probe synthesis cost. , to achieve the effect of real-time monitoring

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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  • Probe composition for detecting three lumen organ tumors
  • Probe composition for detecting three lumen organ tumors
  • Probe composition for detecting three lumen organ tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Embodiment 1

[0081] Such as figure 1 As shown, the implementation process of this application is as follows:

[0082] 1.1.cfDNA extraction and purification

[0083] 1.1.1. Plasma sample preparation:

[0084] 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.

[0085] Table 2

[0086]

[0087]

[0088] 1.1.2. Lysis and conjugation

[0089] 1.1.2.1. Prepare the binding solution / bead mixture according to the table below and mix thoroughly.

[0090] table 3

[0091]

[0092] Add an appropriate volume of plasma sample.

[0093] 1.1.2.2. Thoroughly mix the plasma sample and binding solution / bead mixture.

[0094] 1.1.2.3. Combine cfDNA fully on the rotary mixer for 10 minutes to bind the cfDNA to the magnetic beads.

[0095] 1.1...

Embodiment 2

[0244] 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).

[0245] Table 19

[0246]

[0247]

[0248] Carry out pattern recognition classification and identification on the test samples, firstly judge the methylation level of the pan-cancer specific markers TBX15 and CRYGD genes greater than or equal to 55% and 60%, then preliminarily judge that the sample is a sample with cancer; secondly, judge the methylation level of the gene If the methylation levels of cancer-specific markers OTX1, SFRP2, CDO1, TRIM15, ...

Embodiment 3

[0251] 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).

[0252] Table 20

[0253] 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 SHIS...

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Abstract

The invention discloses a probe composition. According to the composition, through meta analysis of methylation data of a TCGA database and a GEO database, a 4.29 Kbp capture region is screened out, the 4.29 Kbp capture region comprises up to 37 methylation change genes highly related to cancer and non-coding DNA regions, and the method can detect methylation level changes of three kinds of lumen organ cancers such as esophageal cancer, gastric cancer and colorectal cancer at a time. The probe composition can be used for early screening of asymptomatic people and prognosis detection of cancer patients, and the breadth of detection genes of the probe compositino is superior to that of detection genes in the prior art and products.

Description

technical field [0001] This application relates to a cancer gene methylation detection composition, in particular to a probe composition that specifically recognizes DNA sequences treated with bisulfite (Bisulfite), and detection based on high-throughput sequencing (NGS) methods Application of changes in cell-free DNA methylation levels in 3 types of luminal organ tumors including esophageal cancer, gastric cancer and colorectal cancer. 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 cu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12N15/11C12Q1/6886
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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