Model and constructing method thereof

A technology for building methods and identifying models, applied in the fields of biochemical equipment and methods, instruments, biological systems, etc., which can solve the problems of complex subject background, single conclusion, sampling error, etc.

Active Publication Date: 2019-08-09
复旦大学泰州健康科学研究院
View PDF3 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] 1. The current research is basically aimed at the western population, and there are relatively few studies on the oriental population, and the research on metagenomics and cardiovascular diseases in my country is still in its infancy; the genetic background, immune system, living habits and eating habits of the eastern and western people have unique characteristics. For example, in terms of the eating habits of the subjects, the West mainly eats a high-fat and high-calorie diet, while the East mainly eats a carbohydrate diet; in a natural state, the intestinal Presella bacteria content of the Orientals is much lower Higher than Westerners, while Westerners are dominated by Bacteroides content
[0008] 2. Most studies adopt the research idea of ​​sample-control (Case-Control), which lacks the bias control of systematic cohort studies, the background of the subjects is relatively complex, and the investigation of previous diet content and living habits is missing; another On the one hand, the research object is the digestive tract metagenome of patients who have already developed diseases, which is a cross-sectional study, and the correlation analysis between the course of the disease and the bacterial spectrum cannot be carried out through the cohort, so as to judge the early markers, which is of great value in the field of preventive medicine. Limited; again, the number of participants in previous metagenomic studies was small, usually only 20-50 samples, and there was a lack of large-scale studies with hundreds or even more subjects, and sampling errors may exist
[0009] 3. Advanced issues at the technical level
Previous studies mainly rely on the sequencing of the 16s rDNA hypervariable region of bacteria, which can only study the bacterial kingdom in organisms, but can’t do anything about other microorganisms such as viruses, fungi, and parasites; and metagenomic shotgun sequencing with more reliable inspection efficiency Compared with the (Shotgun) method, 16s rDNA sequencing can only know "who is there" in the microbiome, while the latter can also know "what they are doing", which is of great significance for explaining clinical phenotypes and pathogenesis
[0010] 4. The richness of clinical phenotypes of the subjects, especially the lack of cranial imaging data in previous studies, only relying on whether there is atheromatous plaque or infarct lesions, without pathological classification and quantification, is the cause of this problem. single cause

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Model and constructing method thereof
  • Model and constructing method thereof
  • Model and constructing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1 metagenomic DNA purification:

[0050] The metagenomic DNA is extracted and purified, and the specific implementation steps are as follows ( image 3 ).

[0051] 1. Discard the preservation solution in the sampling tube, add the magnetic beads and lysate directly into the sampling tube, shake for 10 minutes, until the bacteria and viruses sticking to the cotton swab fall off into the lysate as much as possible, and transfer the mixed solution to the PowerBead tube of the kit middle.

[0052] 2. Add 60 μl C1 to the above PowerBead tube containing the mixed solution and shake.

[0053] 3. Seal the PowerBead tube with a sealing film, fix it horizontally on the vibrator with thick tape, and adjust it to the maximum vibration speed, and continue to vibrate for 10 minutes.

[0054] 4. Centrifuge at 10000G for 30s at room temperature.

[0055] 5. Take 400μl of the supernatant to a 2ml collection tube.

[0056] 6. Add 250 μl of C2, shake at 4°C for 5 minutes. ...

Embodiment 2

[0069] Example 2 Metagenomic DNA library construction and sequencing ( figure 2 ):

[0070] Using the Nextera XT Shotgun Sequencing Library Construction Kit, the subjects' feces and oral saliva metagenomic DNA was interrupted and library-built, barcodes and mixed samples were marked, and then sequenced on the NovaSeqS2 high-throughput sequencing platform, and a lot of samples were generated The amount of data in 5Mreads / sample.

[0071] The specific steps of metagenomic DNA library construction and sequencing are as follows.

[0072] 1. DNA fragmentation reaction:

[0073] Add 1 ng of purified metagenomic DNA to the reaction system to complete the configuration of the DNA fragmentation reaction system (Table 2).

[0074] Table 2 Configuration list of DNA fragmentation reaction system

[0075]

[0076]

[0077] Place the above reaction system at 55°C, react for 5 minutes (with the hot lid closed), centrifuge briefly until the liquid drops from the tube wall to the bo...

Embodiment 3

[0096] Embodiment 3 metagenomic data analysis:

[0097] Metagenome data analysis, the specific process is as follows:

[0098] Using QIIME2, Silva database, NCBI Blast nr database and MEGAN system for metagenomic sequencing data analysis: quality control, alignment, distance matrix calculation, operational taxonomic unit (OTU) division, diversity analysis, phylogenetic tree construction and signaling pathways And functional annotation of differential flora. The matching database included 660,000 species (25,000 prokaryotic, 84,000 animal, 65,000 plant and 17,000 viral sequences) for class calculation. The calculation process is roughly divided into three parts: quality control and sample pretreatment, substance classification and pedigree tree drawing, functional analysis and differential sample cluster analysis ( Figure 5 ).

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides atherosclerotic early-period identification model construction technology based on metagenome sequencing. According to the technology, an international advance Illumina NovaSeqplatform is utilized. Nextera transposon air gun metagenome sequencing technology is applied on the human feces and oral cavity flora metagenome. In cooperation with traditional high-flux sequencing,metagenome data are generated. Furthermore a database and a system are used for performing metagenome sequencing data analysis. Through performing training set and verifier machine rectification on data, and in cooperation with machine learning and a neural network algorithm, a reliable early-period atherosclerotic identification model is generated; and furthermore verification is performed through expanding a metagenome biomarker. The technology is based on relatively large scale of systematical queue researching. Correlation analysis of pathogenesis change and bacterial spectrum is performedthrough the queue, thereby determining an early-period marker, and supplying a systematical researching method with higher value in a preventative medical science field.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a model and its construction method. Background technique [0002] Cardiovascular disease is the leading cause of death worldwide, of which atherosclerosis accounts for the main pathogenic factor. The analysis results of traditional studies have shown that the risk-related factors of atherosclerosis involve chronic inflammatory diseases such as male sex, age, smoking, hypertension, hyperlipidemia, obesity and diabetes. On the other hand, biomarkers such as elevated circulating high-sensitivity C-reactive protein also suggest atherosclerotic etiology. In recent years, increasing evidence has shown that infections and chronic inflammatory diseases, such as rheumatoid arthritis, are also associated with an increased risk of atherosclerosis. Among them, the inflammatory mechanism caused by bacterial infection is currently considered to be an important cause of atherosclerosis. [0003...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G16B25/00G16B5/00C12Q1/6806C12Q1/6869C12Q1/6883
CPCC12Q1/6883C12Q1/6806C12Q1/6869G16B5/00G16B25/00Y02A90/10
Inventor 陈兴栋朱嗣博庆涛金力
Owner 复旦大学泰州健康科学研究院
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products