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Colorectal cancer microbial marker and application thereof

A colorectal cancer and microorganism technology, applied in the field of microorganisms, can solve problems such as large trauma, high error rate, and low diagnostic value, and achieve the effect of high sensitivity

Inactive Publication Date: 2018-04-13
SUZHOU PRECISION BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, carcinoembryonic antigen (cea) has little diagnostic value for early cases. Although colonoscopy is highly accurate, it is more traumatic. Many high-risk groups refuse to receive regular colonoscopy screening
Studies have shown that mir-92 has the potential to be used as a marker of colorectal cancer, but the error rate of colorectal cancer diagnosis based on the content of mir-92 in blood is as high as 30%.
With the completion of the Human Genome Sequencing Project and the development of high-throughput sequencing technology, genetic screening technology has become a new diagnostic method for colorectal cancer, which has significant advantages in the early diagnosis of colorectal cancer. Rectal cancer has no obvious symptoms in the early stage of the disease, and potential patients still need to be further diagnosed by colonoscopy

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Extraction of Individual Stool Sample DNA

[0041] (1) Fecal samples from 114 subjects (63 colorectal cancer patients and 51 healthy individuals) were collected, frozen immediately, and fused on ice before the experiment;

[0042] (2) Weigh 200 mg of solid feces into a 2 mL centrifuge tube, add 800 μL of fecal DNA extraction buffer (stoolDNA Buffer A), shake and mix well for about 5 min, and centrifuge at 1800 g for 1 min;

[0043] (3) Take out 50 μL of the resuspension in a clean 1.5mL centrifuge tube, add 800μL of Lysis-Binding Buffer (Lysis-Binding Buffer) and vortex to mix, lyse at 70°C for 5min, centrifuge for 5min and transfer the supernatant to a clean 1.5mL centrifuge tube tube;

[0044] (4) Add 20 μL of well-mixed magnetic beads, vortex for 20 seconds, let stand at room temperature for 4 minutes, place on a magnetic rack, let stand for 20 seconds, and absorb the supernatant;

[0045] (5) Add 500 μL of Wash Buffer W1, vortex for 20 s, mix the magnetic ...

Embodiment 2

[0049] Example 2 Sequencing and sequence analysis

[0050] (1) Perform DNA library construction and sequencing, and the off-machine data obtained are called reads;

[0051] (2) Use SOAPdenovo software to assemble the reads to obtain assembled fragments;

[0052] (3) Use MetaGeneMark software to compare the assembled fragments with the reference sequences of microbial markers. According to the similarity between the assembled fragments and the reference sequence of microbial markers is not less than 90%, it is determined that the assembled fragments are derived from microbial markers, and the assembly is obtained through statistics. Fragment abundance;

[0053] (4) Determining the abundance of microbial markers in individual stool samples based on the average abundance of assembled fragments.

Embodiment 3

[0054] Embodiment 3 Determination of Individual State

[0055] Using parallel experiments to determine the relative abundance of microbial markers in stool samples from healthy individuals and patients with colorectal cancer, according to the formula

[0056] cutoff = microbial abundance 结直肠癌 ×0.7+ microbial abundance 健康 ×0.3

[0057] The predetermined critical values ​​shown in Table 1 are obtained. When the abundance of microbial markers in the sample is greater than the critical value, it is diagnosed as a disease state.

[0058] Table 1 Relative abundance of microbial markers in colorectal cancer

[0059] species name

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PUM

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Abstract

The invention provides a colorectal cancer microbial marker and application thereof. The colorectal cancer microbial marker comprises microorganism, wherein the microorganism comprises any one or a combination of at least two of Treptococcus bovis, Clostridium septicum, Parvimonas micra and Helicobacter pylori. The colorectal cancer microbial marker provided by the invention has the potential of being used as a colorectal cancer marker, the content of the colorectal cancer microbial marker in an excrement sample of a patient suffering from colorectal cancer is remarkably higher than the content of the colorectal cancer microbial marker in an excrement sample of a healthy individual, statistical significance is obtained, whether an individual suffers from the colorectal cancer or not can berelatively accurately determined, and a colorectal cancer non-invasively auxiliary diagnosis function is obtained.

Description

technical field [0001] The invention belongs to the field of microorganisms, and relates to a colorectal cancer microbial marker and application thereof. Background technique [0002] Colorectal cancer is one of the most common malignant tumors of the digestive tract. The number of new cases worldwide is about 1 million per year, and the death toll is about 500,000. With the improvement of people's living standards and changes in eating habits, colorectal cancer has become one of the fastest-growing malignant tumors in my country, with about 140,000 new cases each year, accounting for 6.8% of all cancer patients. Tumor ranks 4th. It is predicted that colorectal cancer will become one of the most common malignant tumors in my country, and its morbidity and mortality will continue to rise for a long period of time in the future. Although comprehensive treatment methods such as surgery, chemotherapy, and radiotherapy have been used in the treatment of colorectal cancer, about ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6851C12Q1/6886A61K45/00A61P35/00
CPCA61K45/00C12Q1/6851C12Q1/6886C12Q2535/122C12Q2537/165
Inventor 穆延召朱永亮
Owner SUZHOU PRECISION BIOTECH CO LTD
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