Pathogenic microorganism virulence gene association model as well as establishment method and application thereof

A technology of pathogenic microorganisms and virulence genes, applied in the fields of genomics, bioinformatics, instruments, etc., can solve the problems of easy introduction of pollution, limited detection targets of virulence factor detection, and limited results, so as to achieve the detection of virulence. Comprehensive genes, unlimited sample types, and small sampling requirements

Pending Publication Date: 2021-05-25
GZ VISION GENE TECH CO LTD +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Routine clinical identification of virulence factors requires the cultivation of bacteria first, followed by experimental testing, which is more complicated, the cycle is longer and the operation is easy to introduce pollution
It is also possible to identify virulence factors based on solid-phase chips, but with the method of virulence factor detection kit based on polymerase chain reaction and microfluidic carrier, only a small amount of specific virulence factors can be detected, or a small number of bacterial types, and It is impossible to match the virulence factors with the bacteria. The virulence factors may come from the microbial contamination of the sampled skin or be carried by micro-ecological bacteria, resulting in limited results and false positives
[0008] Therefore, the following problems exist in the detection of virulence factors by traditional methods: the detection method takes a long time, the method is cumbersome to operate, the report result is slow, and the throughput is low. It is often still necessary to identify the strains before the virulence experiment can be carried out, and it is difficult to adapt to the clinic. Treatment needs; for example, the high-virulence Klebsiella pneumoniae often uses the wire drawing test to determine whether it is a high-mucus phenotype (the virulence factor for the high production of capsular polysaccharides formed by the presence of iutA and rmpA or rmpA2 virulence genes at the same time), that is, the need Only when lung culture is positive can the test be judged
However, the detection of virulence factors based on solid-phase chips and microfluidic systems has the characteristics of limited detection targets and high cost, and the corresponding relationship between bacteria and virulence factors cannot be determined.

Method used

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  • Pathogenic microorganism virulence gene association model as well as establishment method and application thereof
  • Pathogenic microorganism virulence gene association model as well as establishment method and application thereof
  • Pathogenic microorganism virulence gene association model as well as establishment method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0074] A method for establishing a pathogenic microorganism virulence gene association model, comprising the following steps:

[0075] 1. Establish a gene database of pathogenic microorganisms.

[0076] Obtaining the reference gene data of pathogenic microorganisms and constructing the pathogenic microorganism gene database can be carried out according to conventional methods. In this example, the pathogenic microorganism genome database constructed by the technical scheme disclosed in the Chinese Invention Patent with the application number of our company is 201910779825.0 for subsequent analysis Comparison.

[0077] It can be understood that the method of the present invention is not limited to specific databases of pathogenic microorganisms, and any open-source and general-purpose databases are acceptable.

[0078] 2. Establish a virulence gene database.

[0079] 2.1 Survey to obtain a list of virulence genes with clear clinical significance.

[0080] Based on the curren...

Embodiment 2

[0113] From the 50,000 clinical samples, 760 samples with Klebsiella pneumoniae detected were selected. First, the sequence ratio of the virulence gene iutA and Klebsiella pneumoniae was calculated, and the log transformation was performed.

[0114] With the iutA-Klebsiella pneumoniae association model of embodiment 1, the sequence ratio of iutA and Klebsiella pneumoniae Klebsiellapneumoniae is divided into 3 classes (A, B, C), such as image 3 As shown, class B is a sample that satisfies the model (derived from Klebsiella pneumoniae), class A may be a sample from Escherichia coli and Shigella flexneri, and class C is a sample without iutA sequence detection;

[0115] Figure 4 It is a box plot of the ratio distribution of the iutA gene and Klebsiella pneumoniae, where the upper and lower dashed lines are the 95% confidence interval of the ratio distribution of the sample that satisfies the model, specifically [5.4e-5,3.7e-3]. In our subsequent application, we can judge wheth...

Embodiment 3

[0118] According to the method of the above-mentioned Example 1, based on the data of 50,000 cases of clinical samples, a variety of pathogenic microorganism virulence gene association models were established, and the following 95% confidence intervals for the ratio of each virulence gene to bacteria were obtained for clinical judgment applications.

[0119] Table 2. 95% confidence intervals for the ratio of virulence genes to bacteria

[0120] virulence gene bacteria 95% lower confidence ratio 95% Upper Confidence Ratio wxya Enteropathogenic Escherichia coli 0.0000182 0.001818652 TcdA Clostridium difficile 0.0000233 0.002330865 TcdB Clostridium difficile 0.0000233 0.002330865 eae Enteropathogenic Escherichia coli 0.0000182 0.001818652 eltA Enterotoxigenic Escherichia coli 0.0000182 0.001818652 eltB Enterotoxigenic Escherichia coli 0.0000182 0.001818652 estIa Enterotoxigenic Escherichia coli 0.0000182 ...

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Abstract

The invention relates to a pathogenic microorganism virulence gene association model and an establishment method and application thereof, and belongs to the technical field of gene detection. The method comprises the following steps: establishing a pathogenic microorganism gene database and a virulence gene database; acquiring pathogenic microorganism metagenome sequencing data of clinical samples to obtain pathogenic microorganisms in each clinical sample and corresponding virulence gene sequence data; carrying out clustering analysis on the sequence data of the pathogenic microorganisms and the sequence data of the virulence genes to obtain a normal distribution model of a single virulence gene and at least one suspected associated pathogenic microorganism, and selecting a model with high pathogenic microorganism abundance and strong correlation between the virulence gene sequence number and the pathogenic microorganism sequence number. The pathogenic microorganism virulence gene correlation model obtained by the method can judge which bacteria each virulence gene comes from, so that the model overcomes the defects of the prior art, and has the advantages of wide application range and high accuracy.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a pathogenic microorganism virulence gene association model and its establishment method and application. Background technique [0002] Virulence factors (VFs) refer to molecules with virulence properties such as invasiveness and toxins produced by the metabolism of bacteria, viruses, and fungi, which indicate the strength of pathogenicity of pathogens. It is mainly used when microorganisms infect the host, colonize in the body, break through the defense barrier of the body, internalize, reproduce and spread, enter and exit the host tissue cells by inhibiting or evading the host's immune response, and obtain nutrition from the host and self-proliferation and growth the goal of. [0003] Bacterial toxins can be divided into two categories: exotoxins and endotoxins according to their sources, properties, and effects. In most cases, exotoxins are generally referred to simpl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G16B20/00G16B50/00
CPCG16B20/00G16B50/00
Inventor 许腾何福生张俊杰曾伟奇李晓蕾苟雪静李永军王小锐苏杭
Owner GZ VISION GENE TECH CO LTD
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