Constipation pathogenic bacteria and application thereof

A technology of bacteria and Shigella, applied in the direction of bacteria, biochemical equipment and methods, microorganisms, etc., to achieve broad industrial prospects and rapid diagnosis

Pending Publication Date: 2022-03-29
陕西安宁云生生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, dysbiosis is likely to be a secondary change in chronic constipation

Method used

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  • Constipation pathogenic bacteria and application thereof
  • Constipation pathogenic bacteria and application thereof
  • Constipation pathogenic bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0216] Embodiment 1, experimental method:

[0217] Specimen and stool culture

[0218]Fresh colon segments were collected from ICC patients undergoing subtotal colectomy plus modified Duhamel procedure at Jinling Hospital (China). Wash the specimen with common solution, and spread the bacterial pellet on the blood plate (1% peptone, 0.3% beef extract, 0.5% NaCl, 1.5% agar and 5% sterile defibrinated sheep blood, pH7.3±0.1 ), and then cultured at 37°C for 12-16 hours. Monoclonal bacteria were inoculated into culture tubes containing 3 ml of LB medium (1% peptone, 0.5% yeast extract and 1% NaCl, pH 7.3±0.1). Culture supernatants were collected and screened.

[0219] Intestinal motility assay and bacterial screening

[0220] Colonic tonometry was performed on bacterial culture supernatants from ICC colons. Briefly, colon segments (approximately 5 mm in length) were isolated from 8-10 week-old C57BL / 6 mice and treated with 2+ HEPES-Tyrode (H-T) buffer (140.6mmol / L NaCl, 2.7m...

Embodiment 2

[0239] Example 2, identification of motility-inhibiting bacteria

[0240] Four colonic biopsies were collected from four patients with refractory chronic constipation, and bacterial monoclonal cultures were performed on blood plates. Then, among the obtained colonies, several thousand single clones were transferred to 3 mL of LB medium without antibiotics, respectively, and cultured at 37°C overnight. The effect of bacterial cultures on intestinal motility was examined. Since the peristaltic contraction frequency of the jejunum is faster relative to the colon, the effect of the culture on the peristalsis of the jejunum was first measured and then confirmed for the colon. After a lot of screening, three colonies with obvious inhibitory effects were obtained, and these bacterial colonies maintained the same inhibitory effect even after passing to the sixth generation. These colonies all showed similar morphology, and their inhibitory activity was also similar, especially the s...

Embodiment 3

[0241] Example 3, Genetic Characterization of PIB

[0242] The PIB 16S ribosomal RNA gene (1464bp) was sequenced. Phylogenetic tree analysis showed that PIB16S rrs was closest to Shigella genus with high bootstrap value (97%). Because the evolutionary distance is not close to Shigella dysenteriae strains ( figure 2 ), so PIB is a novel strain of Shigella. The full genome sequence of PIB contained more non-coding RNA and fewer genes than reported Shigella strains (Table 1). Additionally, the genome size differed between these strains. Therefore, PIB was identified as a novel strain of Shigella.

[0243] Table 1. PIB Genome Characteristics

[0244]

[0245]

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PUM

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Abstract

The invention relates to a novel constipation pathogenic bacterium as well as a screening method and application thereof. The invention further relates to a composition containing the constipation pathogenic bacteria, a reagent for identifying the constipation pathogenic bacteria and application of the composition and the reagent.

Description

technical field [0001] The present invention relates to bacteria and its use, in particular to constipation pathogenic bacteria and its use. Background technique [0002] With changes in diet structure, accelerated pace of life, and the influence of social and psychological factors, chronic constipation has become a chronic disease with high incidence. Among them, the prevalence of chronic constipation in adults in my country is about 4-10%, and the global average prevalence is as high as 16%. Because chronic constipation has a huge impact on the quality of life, physical and mental health, and even life safety of patients, and also greatly increases the social medical burden, the research on chronic constipation has become a major topic of chronic disease research. Although people have conducted extensive basic research and clinical research on chronic constipation, and some important progress has been made, 72.9% of patients are still dissatisfied with the current treatmen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/689C12Q1/04C12N15/11C12R1/01
CPCC12N1/20C12Q1/689C12Q1/04C12Q2600/156
Inventor 朱敏生陈鑫
Owner 陕西安宁云生生物技术有限公司
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