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Method for screening gene marker for intestinal cancer and/or gastric cancer, gene marker screened by the method, and use of gene marker

A technology for gene markers and gastric cancer, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems such as poor specificity and sensitivity, poor user experience, manipulating stool, etc.

Pending Publication Date: 2018-07-20
北京玄鸟飞讯科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Large-scale clinical trials have shown that Cologuard has a better detection rate, but requires feces manipulation and poor user experience; while SEPT9 methylation belongs to liquid biopsy, although the user experience is better, but the specificity and sensitivity are poor (respectively) 80% and 72%)

Method used

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  • Method for screening gene marker for intestinal cancer and/or gastric cancer, gene marker screened by the method, and use of gene marker
  • Method for screening gene marker for intestinal cancer and/or gastric cancer, gene marker screened by the method, and use of gene marker
  • Method for screening gene marker for intestinal cancer and/or gastric cancer, gene marker screened by the method, and use of gene marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Screening of intestinal cancer gene markers

[0031] (1) Extract plasma DNA:

[0032] 10ng of plasma DNA was extracted from samples from 15 patients with intestinal cancer and 18 normal people. This step can be carried out using any methods and reagents suitable for extracting plasma DNA well known to those skilled in the art.

[0033] (2) End-fill the plasma DNA, overhang A and connect with the sequencing adapter:

[0034]According to the Kapa Hyper Perp Kit instructions, prepare a reaction mixture containing 50uL plasma DNA, 7uL End Repair&A-Tailing Buffer and 3uL End Repair&A-Tailing Enzyme mix (total volume is 60uL), incubate at 20°C for 30 minutes, then incubate at 65°C for 30 minutes minute. Prepare the following ligation reaction mixture in a 1.5mL low adsorption EP tube: 5uL Nuclease free water, 30uL Ligation Buffer and 10uL DNA Ligase. Add 5uL of sequencing adapters to 45uL ligation reaction mixture, mix, heat at 20°C for 20 minutes, and then kee...

Embodiment 2

[0053] Example 2. Effectiveness of intestinal cancer gene markers

[0054] This example verifies the effectiveness of the intestinal cancer gene markers of the present invention in detecting intestinal cancer.

[0055] Measure the 5-hmC content of 10 intestinal cancer gene markers of the present invention in the first batch of 59 samples (24 routine intestinal cancers and 35 routine contrasts) according to the method for embodiment 1, and determine the weighting of each genetic marker coefficient.

[0056] The normalized 5-hmC content of each gene marker is multiplied by the corresponding weighting coefficient to obtain the predictor t of the gene marker, and then the predictor t of each gene marker is added to obtain the total predictor T, and then The total predictor T was Logistically transformed according to the following formula to obtain the score P:

[0057]

[0058] If P>0.5, the subject sample has intestinal cancer; if P<0.5, the subject sample is normal.

[005...

Embodiment 3

[0061] Example 3. Screening of gastric cancer gene markers

[0062] The gene markers of gastric cancer were screened according to the method described in Example 1, the only difference being that the samples used were free plasma DNA from 7 gastric cancer patients and 18 normal persons. The screened gastric cancer markers are shown in Table 2.

[0063] Table 2: Average normalized 5-hmC content and weighting coefficients of the gastric cancer gene markers of the present invention

[0064]

[0065]

[0066] As mentioned above, the average normalized 5-hmC content refers to the ratio of the average 5-hmC content of the gene marker in gastric cancer samples to the average 5-hmC content of the same gene marker in normal samples. As can be seen from Table 2, there is a significant difference in the 5-hmC content of the gastric cancer gene markers of the present invention in normal samples and intestinal cancer samples, wherein in gastric cancer samples, three gene markers sho...

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Abstract

The invention relates to a gene marker for detecting the intestinal cancer or the gastric cancer, and a user thereof, and also relates to a method using the gene marker to detect the intestinal canceror the gastric cancer.

Description

technical field [0001] The present invention relates to the field of screening gene markers by high-throughput sequencing. Specifically, the present invention relates to a method for screening colon cancer and / or gastric cancer gene markers by high-throughput sequencing, as well as the gene markers screened by the method and uses thereof. Background technique [0002] With the change of living environment and habits, the incidence of cancer has been increasing year by year in the world in recent years, and people pay more attention to it. Gastric cancer and colon cancer are the second and third most frequently occurring cancers in my country, respectively, and are malignant tumors with very high morbidity and mortality. Based on the existing common early detection and early treatment programs for cancer, the earlier the cancer is detected, the more likely it is to control or even cure cancer early. How to diagnose gastric cancer and intestinal cancer early, treat them in t...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12N15/11C12Q1/68C12Q1/6886C12Q2600/154C12Q2600/158
Inventor 陆星宇宋艳群
Owner 北京玄鸟飞讯科技有限公司
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