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Colorectal cancer biomarker as well as screening method and application thereof

A biomarker and colorectal cancer technology, applied in the field of colorectal cancer biomarkers and its screening, can solve the problems of low penetration rate and complicated preparation process, and achieve high sensitivity and good specificity

Pending Publication Date: 2021-11-30
广西爱生生命科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, colonoscopy is the gold standard for detection of colorectal cancer, but its popularity is very low due to the complicated preparation process and invasive way of detection

Method used

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  • Colorectal cancer biomarker as well as screening method and application thereof
  • Colorectal cancer biomarker as well as screening method and application thereof
  • Colorectal cancer biomarker as well as screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Establishment of intestinal microbial marker model for colorectal cancer

[0057] 1. DNA extraction of intestinal bacteria

[0058] Stool samples from 45 colorectal cancer patients and 46 healthy people were collected for screening intestinal biomarkers of colorectal cancer. The specific steps are as follows:

[0059] 1: Aliquot the above stool samples and freeze them at -80°C;

[0060] 2: Use the Surbiopure Fecal Nucleic Acid Extraction Kit (Guangzhou Saibai Pure Biotechnology Co., Ltd.) to extract the DNA of fecal bacteria from colorectal cancer patients and healthy people described in 1. The specific steps are as follows:

[0061] S1. Get 0.25g stool sample (set a blank control (ddH 2 O) Add the positive control (a nucleic acid with known microbial composition and abundance (ZymoBIOMICS Microbial Community Standard; Cat. No. D6300)) to the Drybeads Tube, add 900 μl S1-Lysis Enhancer, vortex thoroughly, and mix well.

[0062] S2. Add 100 μl of S2-Lysis Enhancer sol...

Embodiment 2

[0102] Methylation detection of ADHFE1 gene in colorectal cancer

[0103] 1. Primer design

[0104] 1. Using ACTB as an internal reference, design primers for fluorescence quantitative detection according to the position of the CpG island of ADHFE1. The position of the CpG island is as follows: figure 1 Shown; ADHFE1 gene detection primers are as follows:

[0105] ADHFE1-PF: 5'-AGGCCTACGGAGCAGTTA-3' (SEQ ID NO.3);

[0106] ADHFE1-PR: 5'-CCACGGAGTAAGAACCAGAAC-3' (SEQ ID NO.4);

[0107] ADHFE1-NF:5'-TTTGGTTTGTTGGTGAGTAGAG-3' (SEQ ID NO.5);

[0108] ADHFE1-NR: 5'-ACACACCAATTAACAACTCCA-3' (SEQ ID NO. 6).

[0109] 2. Extraction of host DNA

[0110] Fecal samples from 45 patients with colorectal cancer and 46 healthy people were collected respectively, and the samples were subpackaged and frozen at -80°C (same as in Example 1). The host DNA of the above samples was extracted with a blood tissue DNA extraction kit (Tiangen Biochemical Technology Co., Ltd.), and the DNA was stor...

Embodiment 3

[0143] Methylation detection of SDC2 gene in colorectal cancer

[0144] 1. Primer design

[0145] 1. Using ACTB as an internal reference, design fluorescent quantitative detection primers according to the position of the CpG island of SDC2. The position of the CpG island is as follows: figure 2 Shown; SDC2 gene detection primers are as follows:

[0146] SDC2-PF: 5'-TCCTCGGAGCTGCCAATC-3' (SEQ ID NO. 7);

[0147] SDC2-PR: 5'-AGGCGGCAGTGTGACTC-3' (SEQ ID NO.8);

[0148] SDC2-NF: 5'-TTTGGTTTGTTGGTGAGTAGAG-3' (SEQ ID NO.9);

[0149] SDC2-NR: 5'-ACACACCAATTAACAACTCCA-3' (SEQ ID NO. 10).

[0150] 2. The extraction, methylation treatment and methylation detection steps of the host DNA are the same as in Example 2.

[0151] 3. Results

[0152] The methylation detection results of SDC2 are as follows: Image 6 Shown: When the positive control primer Tm value=79.5 is used as the critical value, the correct rate of CRC samples is 84.44%, and the correct rate of healthy samples is ...

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Abstract

The invention relates to a colorectal cancer biomarker as well as a screening method and application thereof, and belongs to the technical field of cancer diagnosis. The colorectal cancer biomarker provided by the invention comprises human intestinal microorganisms and / or fecal human target genes; the intestinal microorganisms of the human body comprise one or more than two of Brawtella, Pseudomonas 9, Fecal bacillus, fusiform, Wehonor coccus, Escherichia coli-Shigella, Mycobacterium, Fusobacterium, Porphyrinium, peptostreptococcus and Bacteroides; the fecal human body target spot gene comprises ADHFE1 and / or SDC2. The biomarker combination disclosed by the invention is high in sensitivity and good in specificity, and has a profound prospect for estimating the risk of colorectal cancer. According to the invention, a data model is established through the biomarker, and the model can be used for evaluating the risk of colorectal cancer.

Description

technical field [0001] The invention relates to the technical field of cancer diagnosis, in particular to a colorectal cancer biomarker and its screening method and application. Background technique [0002] Colorectal cancer is one of the most common malignant tumors worldwide, and its incidence ranks third among all malignant tumors. Early prevention is a good strategy to prevent and treat colorectal cancer. Currently, colonoscopy is the gold standard for detecting colorectal cancer, but its popularity is very low due to the complicated preparation process and invasive way of detection. Therefore, it is imminent to develop new ideas for the early prevention and detection of colorectal cancer. Contents of the invention [0003] The purpose of the present invention is to provide a colorectal cancer biomarker and its screening method and application. The biomarker combination of the present invention has high sensitivity and good specificity, and has far-reaching prospec...

Claims

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Application Information

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IPC IPC(8): G16H50/20G16H50/30G16B20/30G06K9/62C12Q1/689C12Q1/6886
CPCG16H50/20G16H50/30G16B20/30C12Q1/6886C12Q1/689C12Q2600/154C12Q2600/158C12Q2600/178G06F18/24323Y02A90/10
Inventor 蒙丽丽芦志龙庞世福陈晓春罗卫飞余秀奇黄连飞甘彩玉王小军叶朋朋
Owner 广西爱生生命科技有限公司
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