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Enteric microorganism combination and application thereof as systemic lupus erythematosus marker

A technology for gut microbes and lupus erythematosus, applied in the field of microbiology, can solve problems such as lack of early detection methods

Active Publication Date: 2021-12-24
梧州市工人医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For systemic lupus erythematosus, there is not yet a very effective early detection method in clinical practice, so it is urgent to establish a highly sensitive and economical method to meet the needs of clinical diagnosis, prevention or treatment of systemic lupus erythematosus

Method used

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  • Enteric microorganism combination and application thereof as systemic lupus erythematosus marker
  • Enteric microorganism combination and application thereof as systemic lupus erythematosus marker
  • Enteric microorganism combination and application thereof as systemic lupus erythematosus marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Study Subjects and Sample Collection

[0075] With informed consent, fecal samples from 300 subjects, including 120 SLE patients and 180 healthy controls, were collected at the Nanfang Hospital of Southern Medical University.

[0076] To be eligible for inclusion in this study, individuals must meet the following criteria for stool sample collection:

[0077] 1) No treatment with antibiotics or immunomodulators, no specific diet (diabetics, vegetarians, etc.) and normal lifestyle (no extra stress) for at least 3 months;

[0078] 2) At least 3 months after any medical intervention;

[0079] 3) No pregnancy, tumor, infection or autoimmune disease or rheumatic disease

[0080] Distribute stool sample collection tubes to the research subjects and inform them of the specific collection method. The subjects take 10-15g of fresh feces and put them in a collection tube with fecal genome protection solution, express them to the hospital laboratory at room temperature...

Embodiment 216

[0081] Example 2 16S rRNA Analysis of Intestinal Microflora

[0082] 2.1 DNA extraction

[0083] DNA extraction was performed using a Fecal Microbial Genome Extraction Kit (Guangdong Nanxin Medical Technology Co., Ltd.) according to the manufacturer's instructions.

[0084] Extracts were treated with DNase-free RNase to eliminate RNA contamination. DNA content was determined using a NanoDrop spectrophotometer, Qubit fluorometer.

[0085] 2.2 DNA library construction and sequencing

[0086] The purified genomic DNA was subjected to two rounds of 16S rRNA gene PCR amplification with bacterial universal primers to complete the construction of the sequencing sample library. 2.0 Fluorometer (Invitrogen, USA) detection concentration, Agilent2100 (Agilent, USA) detection library size; Application Illumina MiSeq (Illumina, San Diego, USA) sequencer is used to sequence the sequence of the amplified 16S rRNA hypervariable region, and the sequencing region is V3-V4 area.

[0087] 2....

Embodiment 3

[0097] Example 3 Clinical detection and verification of intestinal flora in SLE patients

[0098] The above studies have shown that SLE patients are significantly different from healthy people in the classification of intestinal bacteria species, and there are differences in the level of multiple intestinal microorganisms. ).

[0099] According to the correlation between related flora and SLE, SLE can be diagnosed by detecting the abundance of related flora in patient samples.

[0100] Therefore, in this embodiment, 100 feces samples from healthy patients and 100 SLE patients were collected, and the samples were numbered for testing. According to the 16S rRNA gene sequence of the target microorganism, specific fluorescent quantitative PCR primer probes were designed and internal standards were set.

[0101] Table 2 shows the real-time fluorescent quantitative PCR (Quantitative Real-time PCR) specific primer sequences for detecting the abundance of Sporobacter, Butyricimonas,...

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Abstract

The invention discloses an enteric microorganism combination and an application thereof as a systemic lupus erythematosus marker. Particularly, 16s rRNA sequencing is carried out on enteric microorganism samples from systemic lupus erythematosus individuals and healthy individuals to determine Sporobacter, Butyricimonas and Phascolarctobacterium which have level difference in SLE patients, and the relationship between the level difference and the SLE is defined. The enteric microorganisms with the level difference can be used as markers of the SLE.

Description

technical field [0001] The present invention relates to microbiology, in particular to intestinal microbial combination and its application as systemic lupus erythematosus marker. Background technique [0002] Systemic lupus erythematosus (SLE) is a common chronic autoimmune connective tissue disease involving multiple systems. SLE patients are mainly young women, the ratio of women to men is 9:1, and the peak age of onset is 20-40 years old. At present, it is generally believed that its pathogenesis is related to the decrease of T lymphocytes, the decrease of T-reg function, the excessive activation of B cells and the abnormal function. At present, the clinical diagnosis of systemic lupus erythematosus mainly uses clinical symptoms and serological laboratory autoantibodies and other inspection methods, lacking specific auxiliary disease judgment methods and related detection kits. [0003] The gut microbiome is the largest ecosystem in the human body, with more than 10 1...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/689G01N33/569C12Q1/02A61K45/00A61P37/02C12R1/01
CPCC12N1/20C12Q1/689G01N33/56911C12Q1/025A61K45/00A61P37/02C12Q2600/16C12Q2600/136C12Q2600/158C12Q2600/178G01N2800/104G01N2800/52Y02A50/30
Inventor 谢宝钊黄菁梅李水贤赖春桃林智明杨柳陆雪钟金河卢品伶
Owner 梧州市工人医院
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