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Preparation method of high-activity fecal microbiota capsule

A high-activity, fecal bacteria technology, applied in the field of preparation of high-activity fecal bacteria capsules, can solve the problems of patients' clinical improvement, failure to analyze the structure of the bacterial group, etc., and achieve good clinical results

Pending Publication Date: 2019-04-26
广州承葛生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it has been reported to use fecal bacteria freeze-dried powder to prepare fecal bacteria capsules for fecal bacteria transplantation, but they did not analyze the bacterial flora structure in the freeze-dried powder from the overall fecal bacterial flora activity and flora composition, which may be practical for patients. important impact on clinical improvement

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  • Preparation method of high-activity fecal microbiota capsule
  • Preparation method of high-activity fecal microbiota capsule
  • Preparation method of high-activity fecal microbiota capsule

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preparation example Construction

[0032] A preparation method for highly active fecal bacteria capsules, under the protection of acid-resistant enteric-coated capsules, highly active fecal bacteria can reach the patient's intestinal tract to play a role, comprising the following steps:

[0033] S1, preparation of feces liquid;

[0034] S2. Evaluation of fecal liquid activity and flora structure;

[0035] S3. The fecal bacteria liquid obtained in step S2 is added with a freeze-drying protective agent and frozen until the fecal bacteria capsule is transplanted to the patient. When the fecal bacteria capsule is transplanted to the recipient, the fecal bacteria liquid is thawed and centrifuged to obtain the bacteria;

[0036] S4. Dissolving the bacterial cells in step S3 with an appropriate volume of fecal bacteria solvent, and packing the fecal bacterial cells into double-layer acid-resistant enteric-coated capsules.

[0037] In step S1, the preparation method of fecal bacteria liquid is:

[0038] S10. Instrume...

Embodiment 1

[0050] Isolation of fecal bacteria

[0051] S10. Instrument preparation: Seal the separation tank containing the donated feces with a lid with a helical blade stirring rod, and connect the stirring rod to a feces separation machine specially made by Nanjing Famet Company. Connect 5 aseptic and airtight filter tanks to discharge and install them in the separator in sequence. The initial tank is connected to the separation tank with a conduit, and the final tank is connected to the collection tank to form a complete closed system consisting of stirring, filtering and collecting. For separation equipment, the safety cabinet is sterilized by ultraviolet light for 30 minutes before use;

[0052] S11. Start the machine: use the filling pump to fill 500ml of 0.9% normal saline into the separation tank, turn on the mixer at 1000rpm for 5min, then inject 500ml of normal saline, and stir at 1000rpm for 5min; this process completely mixes the feces with the normal saline , so that the i...

Embodiment 2

[0056] Evaluation of Fecal Bacteria Activity During Cryopreservation

[0057] 1) For the bacterial cells obtained in step S13 in Example 1, divide 1 mL of bacterial liquid at five time points of day 0, 1 week, 2 weeks, 3 weeks and 1 month, and place them at -80°C Preservation (except for fresh fecal bacteria);

[0058] 2) Take 10 μl of bacterial solution and mix with 990 μl of 0.9% NaCl solution to obtain a dilution of fecal bacteria;

[0059] 3) Take out the frozen bacterial liquid at different time points, and prepare samples according to the kit Live / Dead BacLightTM BacterialViability and Counting Kit;

[0060] 3) Add 987 μl of 0.9% NaCl solution, 1.5 μl of SYTO9 and 1.5 μl of PI dye to a 2 ml sterilized centrifuge tube, and finally add 10 μl of the fecal bacteria dilution in step 4), mix well and place in the dark Incubation for 15 minutes;

[0061] 4) Setting parameters, counting the ratio of live bacteria by flow cytometry.

[0062] Proportion of viable bacteria duri...

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Abstract

The invention discloses a preparation method of a high-activity fecal microbiota capsule. The high-activity fecal microbiota can reach the intestinal tract of a patient to take effects under the protection of acid-resistant enteric capsules. The preparation method comprises the following steps: S1, preparing fecal microbiota liquid; S2, evaluating the activity and the flora structure of the fecalmicrobiota liquid; S3, adding the fecal microbiota liquid obtained in S2 into a freezing and drying protecting agent, and cryopreserving until the fecal microbiota capsule is transplanted to the patient; when a receptor is transplanted with the fecal microbiota capsule, thawing the fecal microbiota liquid and centrifuging to obtain a thallus; S4, dissolving the thallus in the S3 by using a fecal microbiota solvent with proper volume and loading the fecal microbiota thallus into a double-layer acid-resistant enteric capsule. The preparation method of the high-activity fecal microbiota capsule,disclosed by the invention, has the beneficial effects that the flora has high activity and can be preserved at low temperature for a long time; the method is convenient for implementation of flora transplantation and does not need an invasive implementation mode.

Description

technical field [0001] The invention relates to the technical field of preparation of fecal fungus capsules, in particular to a preparation method of highly active fecal fungus capsules. Background technique [0002] Fecal microbiota transplantation (FMT) is to transplant the functional flora in the feces of healthy people into the gastrointestinal tract of patients, rebuild the intestinal flora with normal functions, and realize the treatment of intestinal and extraintestinal diseases. Before the international cooperation group unified the term, it was translated as "fecal transplantation", "enteric bacteria treatment" or "intestinal microecological transplantation" in a few cases. [0003] The earliest use of feces to treat human diseases was Ge Hong in the Eastern Jin Dynasty (300-400 A.D.). "Squeeze the feces juice, drink it up to one or two liters, it is called Huanglong soup, and it is better if it has been aged for a long time." In 1958, Eiseman et al., University o...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/04C12Q1/04A61K35/74A61K9/48A61P1/00
CPCA61P1/00A61K9/48A61K35/74C12N1/04C12N1/20C12Q1/04
Inventor 陈章然肖传兴杨晓宁姜涛何剑全
Owner 广州承葛生物科技有限公司
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