Method for analyzing intestinal flora and serum metabolites of myositis patient

A technology of intestinal flora and analysis method, applied in the field of biomedicine, can solve problems such as research on intestinal flora of patients with myositis

Pending Publication Date: 2021-09-14
XIANGYA HOSPITAL CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the gut microbiota of patients with myositis has not been systematically studied

Method used

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  • Method for analyzing intestinal flora and serum metabolites of myositis patient
  • Method for analyzing intestinal flora and serum metabolites of myositis patient
  • Method for analyzing intestinal flora and serum metabolites of myositis patient

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The method for analyzing the intestinal flora and serum metabolites of patients with myositis in this example specifically includes performing 16s rRNA sequencing on stool samples from patients with myositis, performing LC-MS / MS analysis on serum samples from patients with myositis, and analyzing the Serum samples from patients with myositis were analyzed by ELISA. ELISA analysis included detecting the level of inflammatory factors in the blood, and then using a Bio-Tek microplate reader to read the OD value, calculating the concentration according to the standard curve, and then using SOD and MDA kits to analyze myositis patients. Changes of oxidative stress indicators SOD and MDA.

[0046] 16s rRNA sequencing includes the following steps:

[0047] S101. Collect stool samples, use Illumina NovaSeq PE250 to perform 16S rRNA gene amplification and sequencing, and obtain raw sequence data;

[0048] S102, analyzing the obtained raw sequence data using qiime2 and R softwar...

Embodiment 2

[0058] On the basis of the aforementioned Example 1, in this example, the control method was used to analyze the intestinal flora and serum metabolites of the normal healthy group and the intestinal flora and serum metabolites of myositis patients.

[0059] Select 10 cases of healthy control group (A: healthy control group) and 10 cases of myositis patients (E: myositis patient group) to participate in the control.

[0060] Blood and stool samples were collected from healthy subjects and patients with myositis. Blood samples were stored in anticoagulant tubes. After centrifugation at 3000 rpm for 10 min, the supernatant serum was transferred into Eppendorf tubes and stored at -80°C for further processing. Fecal samples were collected in fecal collection tubes and stored at -80°C until further processing.

[0061] 1. Perform 16s rRNA sequencing: collect stool samples from healthy controls and myositis patients, and perform sequencing analysis and analysis according to the 16s ...

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Abstract

The invention relates to the technical field of biological medicine and particularly relates to a myositis patient intestinal flora and serum metabolite analysis method. The method comprises steps of conducting 16s rRNA sequencing on an excrement sample of a myositis patient, conducting LC-MS / MS analysis on a serum sample of the myositis patient and conducting ELISA analysis on the serum sample of the myositis patient. ELISA analysis comprises the following steps of detecting the level of inflammatory factors in blood, reading an OD value by using a Bio-Tek microplate reader, calculating the concentration according to a standard curve, and analyzing the change of oxidative stress indexes SOD and MDA of the myositis patient by using an SOD kit and an MDA kit. A problem of incomplete analysis of intestinal colonies and serum of myositis at present is solved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for analyzing intestinal flora and serum metabolites of patients with myositis. Background technique [0002] Idiopathic inflammatory myopathy (IIMs), also known as myositis, is a group of heterogeneous autoimmune diseases whose pathogenesis is still unclear, mainly divided into dermatomyositis (DM), polymyositis (PM) ), polymyositis (PM), necrotizing myopathy (NM) and inclusion body myositis (IBM), myositis is characterized by muscle weakness, skin disease and organ involvement, of which pulmonary involvement is the most common myositis one of the characteristics. Myositis exhibits different clinical and biochemical features, and the response to treatment and prognosis varies greatly. Autoimmunity plays a role in the pathogenesis of myositis. Myositis-specific autoantibodies (myositisspecific autoantibody, MSA) is considered a key biomarker for diagnosis. Myositi...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/30G01N30/32G01N30/34G01N30/72G01N33/48C12Q1/689
CPCG01N30/02G01N30/30G01N30/32G01N30/34G01N30/72G01N33/48C12Q1/689G01N2030/324C12Q2600/158C12Q2600/178
Inventor 徐立群罗月贝罗朝辉杨欢
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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