Microbial marker of liver cirrhosis, and application

A technology of microorganisms and markers, which is applied in the field of microbial markers for liver cirrhosis, and can solve problems such as abnormal plasma membranes of intestinal cells and defects in transport processes

Inactive Publication Date: 2014-12-10
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some studies (Yi JH et al1999, Paik YH et al2003) indicated that it may be due to the overgrowth of intestinal bacteria in patients and the production of endotoxins, inhibiting the protein synthesis of intestinal epithelial cells, morphological changes in intestinal microvilli, abnormal plasma membranes of intestinal cell

Method used

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  • Microbial marker of liver cirrhosis, and application
  • Microbial marker of liver cirrhosis, and application
  • Microbial marker of liver cirrhosis, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Sample Collection and DNA Extraction

[0030] Patients with liver cirrhosis were from the First Affiliated Hospital of Zhejiang University in Hangzhou, and the matched healthy control group was volunteers. A total of 181 stool samples were collected in the experiment, 98 stool samples from Chinese patients with liver cirrhosis and 83 stool samples from healthy Chinese people, each Each individual's fresh feces samples were divided into 200mg / parts, a total of 5 parts, and immediately stored in a -80°C refrigerator.

[0031] Total DNA was extracted from fecal samples of 98 Chinese patients with cirrhosis and 83 healthy Chinese. Phenol trichloromethane treatment to extract DNA method to extract DNA.

Embodiment 2

[0032] Example 2: Construction of library and sequencing

[0033] The DNA library was constructed according to the operating instructions of the instrument manufacturer (Illumina). PE2*100bp sequencing was performed on the library. The library of 181 samples was sequenced on the Illumina HiSeq2000 (Illumina, San Diego, CA) platform. Each sample produced an average of 4.74Gb (sd.±2.04Gb) of high-quality sequencing results, with a total of 858Gb of sequencing data.

[0034] refer to figure 1 The experimental procedure for identifying the relevant microbial markers of liver cirrhosis, the omitted steps or details are well known to those skilled in the art, and several important steps are introduced as described in the following examples.

Embodiment 3

[0035] Example 3: Identification of biomarkers

[0036] 3.1 Basic processing of sequencing data

[0037]After obtaining the sequencing data of 181 samples in the first phase, filter them, and perform quality control according to the following standards: a) remove reads with more than 3 N bases; b) remove low-quality (Q20) N50reads; c ) to remove more than 10 low-quality (Q2) bases or specify the number of tail N bases. Sequences with missing paired reads were considered as single reads for assembly.

[0038] 3.2 Obtaining a cirrhosis microbiome gene set

[0039] The main body of metagenomic biomarkers is genes and corresponding functions, so it is necessary to assemble sequenced sequences and predict genes, remove redundancy, and construct a non-redundant reference gene set. All sample reads were assembled into contigs (assembled fragments) using SOAPdenovo software. The unassembled reads of the samples were combined for de novo assembly. Finally, 4.4 million contigs were...

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Abstract

The invention discloses a microbial marker of liver cirrhosis, and an application. The microbial marker is one or more selected from one microorganism of Veillonella atypica ACS-134-V-Col7a enriched in a patient group suffering the liver cirrhosis or two microorganisms of Bacteroides uniformis ATCC8492 and Clostridiales enriched in a healthy group. The invention also relates to a drug for treating the liver cirrhosis. A drug for treating the liver cirrhosis can promote or increase one or two selected from the two microorganisms of Bacteroides uniformis ATCC8492 and Clostridiales enriched in the healthy group, and/or suppress or clear the microorganism of Veillonella atypica ACS-134-V-Col7a enriched in the patient group suffering the liver cirrhosis. The method detects relative abundance of the microbial marker in intestinal flora and compares the obtained relative abundance value and a predetermined cutoff value. The invention also relates to a beneficial microorganism combination or functional food. The beneficial microorganism combination or functional food comprises one or two of the two microorganisms of Bacteroides uniformis ATCC8492 and Clostridiales enriched in the healthy group. The invention also relates to a kit for detecting the liver cirrhosis, monitoring a treatment process, or producing and screening the drug. The kit is used for detecting the microbial marker.

Description

technical field [0001] The invention relates to the fields of genetic engineering and biomedicine, in particular to a microbial marker of liver cirrhosis and its application. Background technique [0002] Liver cirrhosis (Liver cirrhosis) is a common clinical chronic progressive liver disease, which is diffuse liver damage caused by long-term or repeated effects of one or more etiologies. Liver cirrhosis is an advanced disease of many liver diseases. It is caused by various reasons such as viral hepatitis, alcoholism, nutritional disorders, cholestasis, schistosomiasis, and circulatory disorders. It is characterized by degeneration and necrosis of liver cells. Early liver cirrhosis can be reversed or not progressed through timely prevention and treatment, while advanced liver cirrhosis will be irreversible, seriously affecting the quality of life of patients, and even life-threatening. [0003] The most common cause in China is viral hepatitis, mainly viral hepatitis B, fol...

Claims

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Application Information

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IPC IPC(8): C12N15/31C12Q1/68A23L1/29A61K45/00A61P1/16A23L33/00
Inventor 李兰娟秦楠
Owner ZHEJIANG UNIV
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