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Intestinal anaerobic microorganism culture bottle and preparation method thereof

A technology of anaerobic microorganisms and culture bottles, which is applied in the field of intestinal anaerobic microorganism culture bottles and its preparation, can solve the problems of inconvenient operation and low cost-effectiveness of operation, and achieve convenient purchase and use, simple operation, storage and transportation. The effect of facilitating the work of scientific researchers

Active Publication Date: 2020-07-03
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For long-distance transportation, this dry powder treatment method is very suitable, but for short-distance transportation, the cost-effective operation is not high. From one laboratory to another, the transportation of anaerobic bacteria can only rely on anaerobic bags. And the transfer of the anaerobic tank is not very convenient

Method used

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  • Intestinal anaerobic microorganism culture bottle and preparation method thereof
  • Intestinal anaerobic microorganism culture bottle and preparation method thereof
  • Intestinal anaerobic microorganism culture bottle and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Adopt the method of the present application to prepare several intestinal anaerobic microorganism culture bottles, the specific preparation method is as follows:

[0025] A preparation method of an intestinal anaerobic microorganism culture bottle, the specific preparation method is as follows:

[0026] S1, dissolving powdered RAM basal medium (Haibo) in sterile water at a ratio of 71:1000 to prepare a basal medium solution; dissolving powdered resazurin in sterile water to prepare 0.1wt.% Resazurin aqueous solution; dissolving powdered L-cysteine ​​salt in sterile water to prepare 2.5wt.% L-cysteine ​​salt aqueous solution;

[0027] S2. The mixed solution obtained by mixing 100mL of basal medium solution, 4mL of L-cysteine ​​saline solution and 0.1mL of resazurin aqueous solution was filled into 10 10mL vials respectively, and passed through Inserting a rubber stopper at the bottle mouth of the vial, sealing the hollow aluminum cover at the end of the rubber stopper a...

Embodiment 2

[0034] The 40 bacterial strains in Table 1 below were anaerobically cultured using two intestinal anaerobic microbial culture bottles prepared in Example 1 and Comparative Example 1. The specific cultivation method is as follows: quickly dissolve the freeze-dried powders of 40 kinds of intestinal anaerobic microorganisms in Table 1 in 1 mL of PBS according to the serial number, and use a 1 mL syringe to draw 500 microliters of the dissolved bacteria solution, and inject it into the In the anaerobic microbial culture bottles marked with the same serial number made by Example 1 and Comparative Example 1; all the culture bottles were cultivated in a 37°C incubator for 24 hours, and the turbidity of the cultured microorganisms was measured respectively. The turbidity unit is MCF, 1MCF =3*10^8CFU / mL, CFU is the unit of strain; the turbidity of all culture bottles before cultivation is 0MCF.

[0035] Table 1:

[0036]

[0037]

[0038] As can be seen from the test results in ...

Embodiment 3

[0040] Among the samples that have been cultured in Example 2, take out 40 culture bottles prepared in Example 1 and place them in a refrigerator at 4°C. After 30 days, take them out, mix them, and then rest for 1 hour to return to room temperature; 500 microliters of liquid were drawn from the 40 cultures, injected into 40 new anaerobic microorganism culture bottles prepared in Example 1, and the 40 new culture bottles were placed in a 37°C incubator for 24 hours.

[0041] After the cultivation, the turbidity of the cultured microorganisms in 40 new culture flasks was measured respectively. The test results show that: except Bacteroides PAC002443, Lachnospiraceae PAC001085_s, LachnospiraceaePAC001459_s and Lachnospiraceae PAC001772_s four strains of bacteria did not grow up, the other 36 kinds of bacteria were activated, and the activation rate reached 90%. storage requirements.

[0042] In addition, since the liquid in the intestinal anaerobic microorganism culture bottle o...

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Abstract

The invention discloses an intestinal anaerobic microorganism culture bottle and a preparation method thereof. The culture bottle comprises a culture bottle sealed and packaged and a culture solutionfilled in the culture bottle. The prepration method comprises the steps of sequentially mixing a basic culture medium solution, an L-cysteine saline solution and a resazurin aqueous solution in proportion to obtain a basic culture solution, filling the culture bottle with the basic culture solution, and sealing and packaging the culture bottle; performing high-pressure sterilization on the sealedculture bottle; and finally, mixing a 1640 culture medium solution, BI serum, vitamin K and hemin in proportion to obtain an active additive mixed solution, injecting the active additive mixed solution into the culture bottle which is subjected to high-pressure sterilization and cooled to room temperature, thereby completing the preparation. According to the anaerobic microorganism culture bottle,a culture bottle body and a packaging piece are adopted, all components for preparing the culture solution are commercially available products, the raw materials are easy to obtain, the threshold andcost of anaerobic culture are reduced, anaerobic culture, storage and transportation are easier to operate, the preparation method is simple and easy to implement, the requirement for large-scale production is met, and the use requirements of hospitals and scientific research institutions are met.

Description

technical field [0001] The invention relates to the technical field of microorganism culture, in particular to an intestinal anaerobic microorganism culture bottle and a preparation method thereof. Background technique [0002] At present, most of the microorganisms in the environment cannot be cultured, and the cultureable microorganisms account for about 1% of the total number. The reason why they cannot be cultured is not only the low abundance, but also the insufficient optimization of the culture conditions, including temperature, pH, Oxygen availability, nutrition, etc. Technically, the temperature can be adjusted, and the pH can also be adjusted. The general difficulty lies in anaerobic treatment and nutrition; therefore, the focus of anaerobic microorganism culture in vitro is the simulation of anaerobic environment and its nutritional environment. [0003] At present, the anaerobic culture technology commonly used in research uses anaerobic boxes, anaerobic bags, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/24C12N1/20C12N1/04
CPCC12M23/20C12M23/08C12M23/38C12N1/20C12N1/04
Inventor 周开臣杨柳王荃姚智
Owner TIANJIN MEDICAL UNIV
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