Probe composition for detecting three substantive organ tumors

A composition and probe technology, applied in the field of DNA sequence probe composition, can solve the problems of changing cell development tendency, tumor phenotype, and cancer occurrence and development effects, so as to overcome tumor heterogeneity and save probe synthesis cost. , the effect of prolonging survival

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Embodiment 1

[0082] Such as figure 1 As shown, the implementation process of this application is as follows:

[0083] 1.1.cfDNA extraction and purification

[0084] 1.1.1. Plasma sample preparation:

[0085] 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.

[0086] Table 2

[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]

[0093] Add an appropriate volume of plasma sample.

[0094] 1.1.2.2. Thoroughly mix the plasma sample and binding solution / bead mixture.

[0095] 1.1.2.3. Combine cfDNA fully on the rotary mixer for 10 minutes to bind the cfDNA to the magnetic beads.

[0096] 1.1...

Embodiment 2

[0244] A case of lung cancer sample was detected by the Panel of the present application, and peripheral blood was collected according to the method of Example 1; the library was built, and sequenced by the Illumina platform; the sequencing data was analyzed through the above-mentioned biological information analysis process, and the methylation level was obtained. The results are shown in Table 19 As shown (Table 19 shows the detected target regions greater than or equal to the methylation threshold).

[0245] Table 19

[0246] 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 FMO3 1 171059887 171059936 0.33 Seq ID No.71 FMO3 1 171060010 171060059 0.33 Seq ID No.72 ITIH3 3 52828746 52828819 0.25 Seq ID No.76 CHST12 7 2473529 2473578 0.51 Seq ID No.91 CHST12 7 2473811 2473...

Embodiment 3

[0250] A case of liver cancer sample was detected by the Panel of the present application, and peripheral blood was collected according to the method of Example 1; the library was built, and sequenced by the Illumina platform; the sequencing data was analyzed through the above-mentioned biological information analysis process, and the methylation level was obtained. The results are shown in Table 20 (Table 20 shows the detected target regions greater than or equal to the methylation threshold).

[0251] Table 20

[0252] 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 MTHFD2 2 74425523 74425572 0.6 Seq ID No.73 GLI2 2 121570226 121570275 0.43 Seq ID No.74 RASSF1 3 50378359 50378432 0.54 Seq ID No.15 APCs 5 112073300 112073439 0.37 Seq ID No.23 TRIM15 6 30131001 30131050 ...

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Abstract

Disclosed herein is a probe composition. According to the composition, through meta analysis of methylation data of a TCGA database and a GEO database, a 3.75 Kbp capture area is screened out, the 3.75 Kbp capture area comprises as many as 36 methylation change genes highly related to cancer and a non-coding DNA area, and the method can detect methylation level changes of three substantive organ tumors such as lung cancer, liver cancer and pancreatic 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 composition is superior to that of detection genes in the prior art and products.

Description

technical field [0001] This article relates to a cancer gene methylation detection composition, in particular to a probe composition that specifically recognizes bisulfite-treated DNA sequences, and the detection of lung cancer based on high-throughput sequencing (NGS) methods The application of the changes of cell-free DNA methylation level in 3 kinds of solid organ tumors, including , liver cancer and pancreatic 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 current clinical ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12N15/11C12Q1/68C12Q1/6886
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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