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Method for culturing streptococcus penumoniae rich in teichoic acid

A technique for Streptococcus pneumoniae and a culturing method, which is applied in the field of Streptococcus pneumoniae cultivation, can solve the problems of high value of anti-capsular polysaccharide antibody, interfere with quantitative determination, etc., and achieve easy mass production, good growth state, and teichoic acid content high effect

Active Publication Date: 2011-06-15
LANZHOU INST OF BIOLOGICAL PROD
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0010] In the evaluation of the efficacy of capsular polysaccharide vaccines, the presence of anti-C polysaccharide antibodies in the human body often interferes with the quantitative determination of capsular polysaccharide antibodies. For example, the value of anti-capsular polysaccharide antibodies measured by ELISA is too high

Method used

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  • Method for culturing streptococcus penumoniae rich in teichoic acid
  • Method for culturing streptococcus penumoniae rich in teichoic acid
  • Method for culturing streptococcus penumoniae rich in teichoic acid

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Experimental program
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Effect test

Embodiment 1

[0055] Embodiment 1, the identification of CSR SCS2 Clone1 bacterial strain 1.1 bacterial species identification

[0056] 1.1 Morphological observation

[0057] Use an inoculation loop to scrape the CSR SCS2 clone1 colonies that grow well on the blood agar medium on the glass slide, add a drop of saline, spread evenly with the inoculation loop, and fix it with slight heat on the gas lamp; or add a drop of liquid cultured CSR SCS2 clone1 Bacterial solution was placed on glass slides and fixed with slight heat on a gas lamp. Gram staining was used for staining, and bacterial morphology was observed under an optical microscope (oil lens, 100x).

[0058] The CSR SCS2 clone 1 strain grows well on the blood agar medium. The bacteria are small and spherical, and the colonies are gray and flat, surrounded by a grass-green a hemolytic ring. Most of the bacteria cultured on the solid blood agar slope are arranged in pairs. Most of the cultured bacteria are arranged in short chains.

...

Embodiment 2

[0080] Example 2, Optimization of CSR SCS2 Clone1 Expanded Culture Conditions

[0081] 2.1 Selection of medium

[0082] (1) Streptococcus pneumoniae cultured in compound medium

[0083] The first generation of bacteria was first inoculated on the surface of the solid medium-sheep blood agar medium, and kept at a constant temperature of 37°C in CO 2 After culturing in the incubator for eight hours, use the inoculation loop to scrape the colonies for subculture, re-inoculate on the surface of new solid medium, and streak; 37°C constant temperature CO 2 After culturing in the incubator for 8 hours, use the inoculation loop to scrape the colonies and inoculate them into 200ml liquid medium in a small Erlenmeyer flask, add 50% glucose to make the final concentration to 1% glucose; 2 After cultivating in the incubator for 8 hours, inoculate the bacterial solution in the small triangular flask into the 600ml liquid medium of the large triangular flask, and add 50% glucose to make t...

Embodiment 3

[0094] The optimal concentration of choline chloride in embodiment 3, culture medium

[0095]Choline chloride was added to the liquid medium at a final concentration of 0 μg / ml, 2.5 μg / ml, 5 μg / ml, 20 μg / ml, 40 μg / ml, 80 μg / ml, 150 μg / ml, and 200 μg / ml. The specific operation is the same as that of 2.1 self-made medium cultivation, the only difference is that after the Streptococcus pneumoniae CSR SCS clone1 is inoculated into the large vertical flask, it is shaken in a constant temperature incubation room at 37°C for 6 hours. A small amount of the cultured bacterial solution was taken, and sterilized by adding formaldehyde with a final concentration of 1% for 3 hours. Use a spectrophotometer to detect the OD value of the harvested bacterial solution at 600 nm, see Table 4 for details.

[0096] Harvest the cells by centrifugation, dissolve the cells in an appropriate amount of normal saline, and measure the OD value at 600nm. Dilute the bacterial solution with physiological ...

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Abstract

The invention relates to a method for culturing streptococcus penumoniae rich in teichoic acid, which is characterized in that choline chloride is added in a liquid culture medium, the culture process is optimized, and the obtained streptococcus penumoniae thallus is rich in teichoic acid and is especially suitable to serve as material for extracting streptococcus penumoniae C polysaccharide.

Description

technical field [0001] The invention relates to the cultivation of pathogenic microorganisms, in particular to the cultivation method of Streptococcus pneumoniae. Background technique [0002] Streptococcus pneumoniae (S.pneumoniae), belonging to the genus Streptococcus of the family Bacillus, Lactobacillus, and Streptococcus, is a Gram-positive coccus. It can also be single or short chain in sputum, pus, and lung tissue lesions. Streptococcus pneumoniae has no flagella, no spores, and can form capsules in the body or in serum-containing medium, and the capsules need special staining to be visible. [0003] Streptococcus pneumoniae often lives in the nasopharyngeal cavity of normal people. Most of them are not pathogenic or have weak pathogenicity, and only a few have pathogenicity. As an opportunistic pathogen, Streptococcus pneumoniae usually forms a carrier state. When the body's immunity is weakened, especially after respiratory virus infection or infants, elderly and ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/46
Inventor 沈荣任珍芸
Owner LANZHOU INST OF BIOLOGICAL PROD
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