Intestinal flora maker for intestinal cancer and application of intestinal flora maker

A technology of intestinal flora and markers, applied in the field of microbiology and genetic engineering, can solve problems such as low survival rate and lost treatment opportunity, and achieve good predictive effect

Inactive Publication Date: 2019-11-05
FUJIAN HEALTH COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Colorectal cancer is so common, but most of the patients are found in the late stage and lost the best opportunity for treatment, resulting in a very low 5-year survival rate for patients with colorectal cancer after diagnosis; in my country, one person dies of colorectal cancer every 5 minutes

Method used

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  • Intestinal flora maker for intestinal cancer and application of intestinal flora maker
  • Intestinal flora maker for intestinal cancer and application of intestinal flora maker
  • Intestinal flora maker for intestinal cancer and application of intestinal flora maker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Construction of the 16S rRNA gene library of the intestinal flora

[0026] 1 sample source

[0027] 101 fresh stool samples from healthy people and 65 fresh stool samples from patients with bowel cancer.

[0028] 2 Extraction of DNA from stool samples

[0029] DNA was extracted from stool samples by spin column method.

[0030] (1) Column balance: add 500uL of equilibration buffer EQ (Buffer EQ) to the adsorption column MP1 (the adsorption column is placed in the collection tube), centrifuge at 12,000rpm (~13,400×g) for 1min, and discard the waste in the collection tube. liquid, put the adsorption column back into the collection tube.

[0031] (2) Weigh 180-220mg of stool sample into a 2ml centrifuge tube, and place the tube on ice.

[0032] (3) Add 1.2ml buffer GSL to the sample, shake intermittently for 1min until the sample is well mixed.

[0033] (4) Incubate at 70°C for 10 minutes.

[0034] (5) Vortex for 15 sec and centrifuge at 12,000 rpm (~13,400...

Embodiment 2

[0099] The sequencing of embodiment 2 library

[0100] 1 Sequencing steps

[0101] 1.1 Library Mixing

[0102] Mix the libraries that need to be sequenced according to the number of 30,000-50,000 tags to be tested;

[0103] 1.2 Library Quality Inspection

[0104] The mixed sample was subjected to Qubit3.0 and Agilent 2200Bioanalyzer quality inspection;

[0105] 1.3 Library Dilution

[0106] Dilute the library to the upper concentration with Hyb Buffer;

[0107] 1.4 Library Denaturation

[0108] Use 0.2N NaOH to denature the library for 5 minutes. After denaturation, you can choose 0.2M Tris-HCl pH 7.0 to neutralize the library after denaturation;

[0109] 1.5 Denaturing dilution

[0110] Dilute the denatured library with Hyb Buffer;

[0111] 1.6 Machine dilution

[0112] Finally, use Hyb Buffer to perform final dilution on the denatured dilution library;

[0113] 1.7 On-machine sequencing

[0114] Dilute the library on the machine, put it into the reagent tank, put ...

Embodiment 3

[0117] Example 3 Analysis of sequencing data

[0118] 1 Sequencing data splicing

[0119] According to the Barcode sequence, the data of each sample is split from the off-machine data, and after the Barcode is cut off, the reads of each sample are spliced ​​using FLASH. The spliced ​​sequence obtained is the original Tags data (RawTags); The filtering process to obtain high-quality Tags data (Clean Tags). Referring to QIIME's Tags quality control process, perform the following operations: a) Tags interception: Raw Tags from continuous low-quality values ​​(default quality threshold is <= 19) bases to the set length (default length is 3) of the first A low-quality base site truncation; b) Tags length filtering: the Tags data set obtained after the Tags are truncated, further filters out the Tags in which the length of continuous high-quality bases is less than 75% of the Tags length. The Tags obtained after the above treatment remove primers and chimeras.

[0120] 2 Sequenci...

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Abstract

The invention discloses an intestinal flora maker for intestinal cancer and an application of the intestinal flora maker, and belongs to the technical fields of microorganisms and genetic engineering.The intestinal flora maker comprises megasphaera, veillenella, streptococci, bilophila sp., haemophilus, corynebacterium or Escherichia-Shigella. Through detection by the intestinal flora maker, risks of individuals suffering from intestinal cancer can be assessed, and auxiliary reference can be provided for disease diagnosis and prognosis monitoring.

Description

technical field [0001] The invention belongs to the technical field of microorganisms and genetic engineering, and in particular relates to a marker of intestinal flora and its application in the detection of intestinal flora of intestinal cancer, the prediction of the risk of intestinal cancer, and the monitoring of the prognosis of intestinal cancer. Background technique [0002] Colon cancer is a common malignant disease of the digestive system, which seriously threatens human health. Colon cancer has become the most common malignant tumor affecting the health of Chinese people. Colorectal cancer is so common, but most of the patients are found in the advanced stage and lost the best opportunity for treatment, resulting in a very low 5-year survival rate of patients with colorectal cancer after diagnosis; in my country, one person dies of colorectal cancer every 5 minutes . Studies have shown that bowel carcinogenesis is a multifactorial process in which a series of diff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/689C12Q1/04
CPCC12Q1/6886C12Q1/689C12Q2600/136C12Q2600/118
Inventor 李泳宁彭臻菲魏碧娜宋光运
Owner FUJIAN HEALTH COLLEGE
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