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In-vitro simulated culture method for enteric microorganisms

A technology of gut microbes and in vitro simulation, applied in the field of in vitro simulation culture of gut microbes, can solve the problems of large individual differences and distinctions of gut microbes

Active Publication Date: 2018-06-05
湖南海路生物科技有限公司 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

On the other hand, due to the huge individual differences in gut microbes, it is difficult to distinguish which are caused by individual differences and which are caused by diseases from the numerous small and non-significant differences

Method used

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  • In-vitro simulated culture method for enteric microorganisms
  • In-vitro simulated culture method for enteric microorganisms
  • In-vitro simulated culture method for enteric microorganisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Response value of intestinal microorganisms to specific oligopolysaccharides through in vitro batch fermentation-healthy adults

[0043] 1. Culture medium preparation

[0044] The formula of YCFA basal medium is as follows: 10g / L typtone, 2.5g / L yeast extract, 0.8g / L L-cysteine ​​hydrochloride, 0.05g / L heme, 4.5g / L NaCl, 4.5g / L CaCl 2 ·6H 2 O 0.09g / L, KH 2 PO 4 0.45g / L, K 2 HPO 4 0.45g / L, MgSO 4 ·7H 2 O 0.09g / L, trace elements, solvent is deionized water, pH value is natural. The final concentration of trace elements in the YCFA basal medium was as follows: resazurin 0.1 mg / L, biotin 10 μg / L, cobalamin 10 μg / L, p-aminobenzoic acid 30 μg / L, folic acid 50 μg / L, pyridoxine Amine 150μg / L, thiamine 50μg / L, riboflavin 50μg / L.

[0045] The YCFA-oligosaccharide medium is to add oligosaccharides or sugar alcohols with a final concentration of 8g / L to the YCFA basal medium. Oligosaccharides refer to polymers of single or multiple monosaccharides linked by α-1...

Embodiment 2

[0112] Example 2 In vitro fermentation of intestinal microorganisms reveals that there is an imbalance of intestinal microorganisms in IBS patients. .

[0113] In this embodiment, the content of metabolites in the feces of the IBS group and the healthy group and their in vitro fermentation liquids were mainly detected. It was found that although there was no difference between the two groups in SCFA in feces, after in vitro fermentation, the SCFA response value of the IBS group to the fermentation of specific oligopolysaccharides was significantly different from that of the healthy group. At present, there is no clinical diagnosis method for IBS in the medical field, and the disease can only be judged by scoring questionnaires. There is no clear conclusion on the relationship between IBS and intestinal flora. The in vitro fermentation involved in the present invention can distinguish IBS from the normal group through the metabolites of in vitro fermented microorganisms, the d...

Embodiment 3

[0137] Example 3 The in vitro fermentation of intestinal microorganisms reveals that there is a certain degree of imbalance in the intestinal microecology of patients with constipation.

[0138] In this embodiment, the content of metabolites short-chain fatty acids (SCFA) in the feces of the constipation group and the healthy group and their in vitro fermentation broth were mainly detected. It was found that although there was no difference between the two groups of SCFA in feces, after in vitro fermentation, the SCFA response value of the constipation group to the fermentation of specific oligopolysaccharides was significantly different from that of the healthy group.

[0139] 1. Preparation of stool samples

[0140] Human stool samples were provided by 14 healthy volunteers (healthy group) and 14 constipation patients (constipated group). Volunteers were required to be in good health and had not taken antibiotics or prebiotics within one month before sampling. The preparat...

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Abstract

The invention discloses an in-vitro simulated culture method for enteric microorganisms. The method comprises the steps of inoculating an excrement sample into a fermentation culture medium, carryingout anaerobic fermentation culture at 37 DEG C, so as to obtain fermentation liquid containing enteric microorganisms, wherein the fermentation culture medium is a YCFA basic culture medium containing6g / L-10g / L of carbohydrate, and carbohydrate is polysaccharide, oligosaccharide or sugar alcohol. By detecting a series respond values of the enteric microorganisms to a specific carbon source in anin-vitro simulated fermentation system, the metabolic capability of the enteric microorganisms and the level of probiotics are directly reflected, and direct evaluation of the functions of a complex ecosystem, namely the enteric microorganisms, is realized.

Description

(1) Technical field [0001] The invention relates to an in vitro simulation culture method for intestinal microorganisms. (2) Background technology [0002] With the rapid development of molecular biology technology, especially the completion of human genome sequencing technology and human intestinal microbial 16S rRNA gene sequencing, we have a deeper understanding of the factors that affect human health. Gut microbes are another factor that is very closely related to health besides processing our own genes. At present, it has been proved that intestinal microorganisms affect the host's conversion efficiency of energy in food, affect the host's weight, and participate in the pathogenesis of metabolic syndrome, fatty liver, chronic colitis, and intestinal cancer. Even, intestinal bacteria are involved in the metabolism of serotonin, which affects the mental state of the host. Therefore, the concept of human health has a new meaning. In addition to the normal function of its...

Claims

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Application Information

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IPC IPC(8): C12N1/00
CPCC12N1/00
Inventor 朱立颖王欣刘伟皮雄娥费迪波陈镜刚李降龙
Owner 湖南海路生物科技有限公司
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