Microbial marker related to colorectal cancer and application thereof

A colorectal cancer and microorganism technology, applied in the field of microorganisms, can solve the problems of intolerance or refusal of screening for colorectal cancer risk groups, the high price of detection technology, and the suffering of the subjects, and achieves good application prospects and practical significance. Accuracy and safety, high sensitivity effects

Inactive Publication Date: 2021-02-26
天津奇云诺德生物医学有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 1985, the world's first bowel cancer screening technology appeared: rigid sigmoidoscopy. After decades of development, colonoscopy technology has continued to improve. Although this technology is the gold standard for bowel cancer screening, it is a more accurate screening method. However, the invasiveness and bowel preparation of this technique may cause a certain degree of distress to the subjects, making it difficult for many people at risk of colorectal cancer to tolerate or refuse screening
The first non-invasive screening technique, the fecal occult blood test (FOBT), was proposed in 1967, and the fecal immunochemical test (FIT) technology was introduced in the 1980s. Although these two technologies have the advantages of non-invasive, rapid, and non-invasive, but low sensitivity and high false positive
In the 1990s, DNA-based bowel cancer screening technology appeared, which has the advantages of non-invasiveness and high sensitivity at the same time, but the detection technology is expensive and cost-effective

Method used

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  • Microbial marker related to colorectal cancer and application thereof
  • Microbial marker related to colorectal cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the extraction of DNA sample.

[0044] (1) Collect individual feces samples, freeze them immediately, and put them on ice before the experiment;

[0045] (2) Weigh 200 mg of fixed feces into a 2 mL centrifuge tube, add 800 μL of fecal DNA extraction buffer, shake and mix well for 5 min, and centrifuge at 1800 g for 1 min;

[0046] (3) Take 50 μL of the suspension into a 1.5 mL centrifuge tube, add 800 μL of lysate, vortex and mix, lyse at 70°C for 5 min, centrifuge for 5 min, and transfer the supernatant to a clean 1.5 mL centrifuge tube;

[0047] (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;

[0048] (5) Add 500 μL of washing solution Ⅰ, vortex for 20 seconds, mix the magnetic beads, place them on a magnetic rack, let them stand for 20 seconds, and discard the supernatant;

[0049] (6) Add 750 μL of washing so...

Embodiment 2

[0053] Example 2, Quantitative Detection of Microbial Markers

[0054] The quantitative detection of microbial markers adopts the Taqman qPCR method, and the probes and primers used are shown in SEQ ID NO.01-09.

[0055] The specific steps of this embodiment will be described below by taking the TaqMan Master Mix kit product of Suzhou Xinhai Biotechnology Co., Ltd. as an example.

[0056] (1) Perform the reaction according to the qPCR reaction system shown in Table 1 to prepare the PCR reaction solution.

[0057] Table 1. qPCR reaction system

[0058] Reagent Dosage concentration Taqman Master Mix (2×) 12.5μL 1× PCR forward primer (10 μM) 1μL 400nM PCR reverse primer (10 μM) 1μL 400nM Probe (0.2uM) 1μL 400nM Sample DNA to be tested 1μL 1-100ng Sterilized water 8.5μL /

[0059] (2) After the PCR reaction solution is prepared, mix it upside down and centrifuge, dispense it into a 96-well PCR reaction plate, ...

Embodiment 3

[0064] Example 3, colorectal cancer risk assessment model.

[0065] The colorectal cancer risk assessment model uses the random forest algorithm to train and test the model using the abundance of three microbial markers from 357 healthy samples and 354 colorectal cancer patient samples, and finally establishes the colorectal cancer risk assessment model, with specific steps as follows:

[0066] Step 1) Using the method described in Example 1 to extract DNA fragments from 357 healthy samples and 354 colorectal cancer samples;

[0067] Step 2) Use as described in Implementation 2 to detect the content of the target gene fragments of the three microbial markers in all samples and the gene content of the internal reference 16S rDNA;

[0068] Step 3) According to the content of the target gene fragment of the microbial marker described in step 2) and the gene content of the internal reference 16S rDNA, calculate the abundance of the corresponding microbial marker;

[0069] Step 4...

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Abstract

The invention provides a microbial marker related to colorectal cancer and an application of the microbial marker. The microbial marker comprises the following components: a microbial marker 1) Peptostreptococcus stomatis; a microbial marker 2) Porphyromonas asaccharolytica; and a microbial marker 3) is Gemella morbillorum. The abundance of the three kinds of bacteria in colorectal cancer patientsis remarkably increased. The invention also provides an application of the biomarker as a detection target in preparation of a detection reagent or a kit and an application in prediction of colorectal cancer risk. The microbial marker disclosed by the invention has the potential of serving as a colorectal cancer marker, is high in accuracy and good in specificity, and has the effect of non-invasive auxiliary diagnosis of colorectal cancer.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a microbial marker related to colorectal cancer and its application. Background technique [0002] Colorectal cancer (Colorectal cancer) is the third most frequently occurring cancer in my country after lung cancer and gastric cancer. With 429,200 cases and 281,000 deaths, the prevention and control situation is severe. The incidence of colorectal cancer is related to age, environment and other factors. About 90% of patients are over 40 years old, but the proportion of young patients with colorectal cancer has gradually increased in recent years, showing a younger trend. The occurrence of colorectal cancer is statistically positively correlated with economic development, indicating that certain reasons in the environment of human economic civilization may lead to the occurrence of colorectal cancer, including diet, exercise and other factors. Statistics show that about 6% of color...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/689C12Q1/6895C12Q2561/113C12Q2563/107C12Q2545/114C12Q2561/101
Inventor 罗奇斌陈卫华申玉林任毅廖胜光
Owner 天津奇云诺德生物医学有限公司
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