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Diarrhea-type irritable bowel syndrome intestinal microbial marker and application thereof

A technology for irritable bowel syndrome and gut microbes, applied in the field of microbiology, can solve problems of limited value and achieve weight loss and reduced dietary intake

Pending Publication Date: 2022-02-15
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although intestinal motility, stool properties, autonomic nervous reactivity, and fecal trypsin levels have been used as potential biomarkers for IBS-D, the value of such indicators in diagnosing IBS-D is limited

Method used

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  • Diarrhea-type irritable bowel syndrome intestinal microbial marker and application thereof
  • Diarrhea-type irritable bowel syndrome intestinal microbial marker and application thereof
  • Diarrhea-type irritable bowel syndrome intestinal microbial marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 Chronic bondage combined gavage with senna leaves

[0032] Sixteen SPF-grade C57BL / 6J mice, weighing (20±2) g, male, were provided by the Guangdong Provincial Medical Animal Center, and the animal qualification certificate number: SCXK (Guangdong) 2018-0002. Before the experiment, the animals were placed in the laboratory to adapt to the environment, free to eat and drink, room temperature (22±2)°C, and normal circadian rhythm. During the experiment, the feeding and management of experimental animals and the experimental operation process complied with the relevant regulations of the Experimental Animal Center of Guangdong Pharmaceutical University.

[0033] Weigh 60g of senna leaves and decoct in 300mL of distilled water. After 10 min, the supernatant was obtained by filtering with double gauze. After decocting at high temperature, concentrate the mass concentration to 0.6g / mL, store in a -20°C refrigerator, heat in a water bath at 25°C before administra...

Embodiment 2

[0034] Example 2 Model Evaluation

[0035] The total number of defecation particles and the number of loose stool particles of mice were calculated by filter paper blotting method 6 hours after modeling. The judgment of loose stools is judged by whether there is stain on the filter paper. Loose stool rate (%) = number of loose stools / total number of stools × 100%. The loose stools were graded according to the diameter of the stain on the filter paper after the model was established: grade 1, the diameter of the stain was <1 cm; grade 2, the diameter of the stain was 1-2 cm; grade 3, the diameter of the stain was 2-3 cm ; Grade 4, the diameter of the stain is 3-4cm; Grade 5, the diameter of the stain is 4-5cm. Diarrhea index = loose stool rate × loose stool grade. The mice were fasted for 24 hours, a 6Fr urinary catheter lubricated with paraffin oil was inserted through the anus, the end of the balloon was placed about 1.0 cm away from the anus, and the catheter and the base...

Embodiment 3

[0037] Example 3 Feces collection and detection

[0038] Six healthy IBS-D mice and six healthy mice were selected, and the feces samples were all from the adult male C57 mouse model raised in the SPF animal room. Use sterile tweezers to put the naturally excreted fresh mouse model feces into a sterile dry collection tube, place it in liquid nitrogen, send it back to the laboratory within 30 minutes, and store it in a -80°C refrigerator. By using the PowerSoil DNA IsolationKit kit to extract the microbial genomic DNA in the feces samples of each group of mice, design primers according to the conserved regions, that is, the primers shown in SEQ ID NO.1-2 of this application, add sequencing at the end of the primers Adapter, perform PCR amplification and purify, quantify and homogenize the product Purify, quantify and homogenize (PCR products are quantified according to electrophoresis (ImageJ software) results, after mixing samples according to the mass ratio 1:1, use OMEGA DNA...

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Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a diarrhea-type irritable bowel syndrome intestinal microbial marker and application thereof. The enteric microbe marker comprises a Staphyloccus genus, an Akkermansia genus, an Enterobacter genus, a Dubosiella genus and a Lactobacillus genus. The level change of intestinal flora in mice with diarrhea-type irritable bowel syndrome diseases is discovered for the first time, early diagnosis of diarrhea-type irritable bowel syndrome is realized, and the kit has the advantages of high specificity, high sensitivity, simplicity in collection and treatment of detection objects, no invasion, low cost and the like; and the discovery of the marker provides reference for diagnosing the microbial diarrhea-type irritable bowel syndrome.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to an intestinal microbial marker for diarrhea-type irritable bowel syndrome and an application thereof. Background technique [0002] Diarrhea-predominant irritable bowel syndrome (IBS-D) is a gastrointestinal disordered enteropathy with diarrhea, abdominal distension and abdominal pain as the main symptoms, and often occurs repeatedly. Because the etiology and pathogenesis of IBS-D have not been fully clarified, and the clinical manifestations lack specificity, the diagnosis is difficult. The identification of diagnostic biomarkers in irritable bowel syndrome has been a matter of interest and may lead to rapid diagnosis of IBS-D in the future. At present, the diagnosis of IBS-D is mainly based on clinical symptoms, so the diagnosis is subjective, and it is easy to miss and misdiagnose. With the deepening of basic and clinical research, some potential IBS-relat...

Claims

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Application Information

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IPC IPC(8): C12Q1/06C12Q1/14C12Q1/689C12Q1/6869G16B20/30G16B50/00G06F16/906C12R1/44C12R1/225C12R1/01
CPCC12Q1/06C12Q1/14C12Q1/689C12Q1/6869G16B20/30G16B50/00G06F16/906G01N2333/31G01N2333/335G01N2333/265G01N2333/195G01N2800/065C12Q2600/158C12Q2600/178C12Q2531/113C12Q2535/122
Inventor 褚夫江凌茜文徐雅琴
Owner GUANGDONG PHARMA UNIV
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