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Specific solid streptococcus culture medium, preparation method and application thereof

A solid medium and specific technology, applied in the field of microbiology, can solve the problems that the monosaccharide medium is easy to be contaminated with bacteria, prone to bacterial contamination, and no clear configuration method is given, so that the preparation steps are simple and clear, and the operation method is simple Effect

Pending Publication Date: 2019-07-09
INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE JIANGXI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] (1) At present, the monosaccharide-liquid medium formula provided by Santi does not clearly specify the configuration method, and the monosaccharide-liquid medium is prone to bacterial contamination during the configuration of the medium and the bacterial culture process
[0009] (2) There is no monosaccharide-solid medium production technology suitable for streptococcus culture, so single colony purification cannot be performed by streak culture, and can only be subcultured in liquid medium
[0013] The present invention improves on the basis of the liquid medium formula provided by Santi, provides a preparation method of streptococcus-specific monosaccharide-solid medium, and realizes the research on the purification of streptococcus single colony on the monosaccharide-solid medium demand, and at the same time solve the phenomenon that the monosaccharide medium is easy to be contaminated with bacteria

Method used

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  • Specific solid streptococcus culture medium, preparation method and application thereof
  • Specific solid streptococcus culture medium, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Prepare 1L of 1% glucose-solid medium:

[0051] (1) Weigh 0.2g of nalidixic acid powder, dissolve it in 10mL of 0.1M / L sodium hydroxide solution, filter it through a 0.22μm sterile filter membrane and dispense it into 1.5mL EP tubes, wrap it in tinfoil paper and place in- Store at 20°C.

[0052] (2) Weigh 10g of peptone, 5g of tryptone, 5g of yeast extract, 15g of agar powder and 2.5g of KCl, add 900mL of double distilled water to dissolve, and use an autoclave to sterilize at 121°C for 15 minutes. Cool to 60°C.

[0053] (3) Weigh 0.06g of urea, 0.1742g of arginine and 10g of glucose, add 100mL of double distilled water to fully dissolve, and filter into the culture medium with a 0.22μm sterile filter membrane.

[0054] (4) Add 1 mL of nalidixic acid solution to 1 L of medium, and adjust the pH of the medium to 7.0 with sterile HCl solution.

[0055] (5) Quickly pour the flat plate, and solidify after cooling. Take a piece of culture medium and place it in a constan...

Embodiment 2

[0061] Prepare 500mL of 0.5% sucrose-solid medium:

[0062] (1) Weigh 0.2g of nalidixic acid powder, dissolve it in 10mL of 0.1M / L sodium hydroxide solution, filter it through a 0.22μm sterile filter membrane and dispense it into 1.5mL EP tubes, wrap it in tinfoil paper and place in- Store at 20°C.

[0063] (2) Weigh 5g of peptone, 2.5g of tryptone, 2.5g of yeast extract, 7.5g of agar powder and 1.25g of KCl, add 450mL of double distilled water to dissolve, and use an autoclave to sterilize at 121°C for 15 minutes and cool to 60°C.

[0064] (3) Weigh 0.03g of urea, 0.0871g of arginine and 2.5g of sucrose powder, add 50mL of double distilled water to fully dissolve, and filter into the culture medium with a 0.22μm sterile filter membrane.

[0065] (4) Add 0.5 mL of nalidixic acid solution to 500 mL of medium, and adjust the pH of the medium to 7.0 with sterile HCl solution.

[0066] (5) Quickly pour the flat plate, and solidify after cooling. Take a piece of culture medium ...

Embodiment 3

[0071] Investigate the inhibitory effect of monosaccharide-solid medium in the present invention to Gram-negative bacteria, concrete experimental process is as follows:

[0072] (1) Prepare 1% glucose-solid medium containing nalidixic acid to prepare a plate, see Example 1.

[0073] (2) Prepare 1% glucose-solid medium without adding nalidixic acid: weigh 10g peptone, 5g tryptone, 5g yeast extract, 15g agar powder and 2.5g KCl, add 900mL double distilled water to dissolve , use an autoclave to sterilize at 121°C for 15 minutes, cool to 60°C; weigh 0.06g urea, 0.1742g arginine and 10g glucose, add 100mL double distilled water to dissolve, and filter through a 0.22μm sterile filter membrane Filter into the culture medium; pour the plate quickly, take a piece of culture medium after solidification, and place it in a constant temperature incubator at 37°C for 24 hours to confirm the sterile contamination; put the plate upside down and put it in a dark place at 4°C for later use.

...

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Abstract

The invention belongs to the technical field of microbes and discloses a specific solid streptococcus culture medium, a preparation method and application thereof. The specific solid streptococcus culture medium contains 10 g / L of peptone, 5 g / L of tryptone, 5 g / L of a yeast extract, 15 g / L of an agar powder, 2.5 g / L of KCl, 0.06 g / L of urea, 0.1742 g / L of arginine, 20 mg / L of nalidixic acid and any mass / volume ratio of monosaccharide according to the mass / volume ratio. The nalidixic acid is added into the culture medium so that the problem of bacterial infection easily occurring in the experimental operation process can be solved, and meanwhile the solid culture medium can also be used for screening, identifying, separating and purifying streptococci and the like.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a streptococcus-specific solid medium, a preparation method and an application thereof. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: [0003] Streptococcus, especially conditionally pathogenic Streptococcus, such as Streptococcus pyogenes, Streptococcus viridans, Streptococcus pneumoniae, Streptococcus agalactiae, Streptococcus mutans, Streptococcus suis, etc., colonize the host (human Oropharyngeal cavity, upper respiratory tract, gastrointestinal tract, reproductive tract and other mucosal surfaces maintain a dynamic balance with the host's immune system, which is of great significance for ensuring the health of the host. [0004] The type and abundance of carbohydrates are important environmental factors that affect the balance of flora. Undigested starch and fiber in food, glucose and oligosacc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12Q1/14C12R1/46
CPCC12N1/20C12Q1/14C12Q1/045G01N2333/315
Inventor 谭美芳李海琴曾艳兵康昭风魏岳
Owner INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE JIANGXI ACAD OF AGRI SCI
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