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Avirulent type-2 pig streptococcus strain and its prepn and application

A technology of Streptococcus suis and microbial strains, applied in the field of bacteria, can solve the problems of small inoculation dose, multiple inoculations, poor vaccine protection, etc., and achieve strong protective effect

Inactive Publication Date: 2010-10-27
中国人民解放军南京军区军事医学研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The inactivated vaccine is an inactivated whole-cell dead vaccine. Its biggest advantage is that it is easy to prepare and produce, and it is stable and easy to store; but the disadvantage is that it needs multiple vaccinations, large vaccination doses, local or systemic toxicity and side effects The reaction is obvious, and some vaccines have poor protection
The advantage of the live attenuated vaccine is that it only needs to be vaccinated once, the inoculation dose is small, the immune effect is better, and the immunity is long-lasting; the disadvantage is that it is unstable, difficult to store, and virulence may reverse mutation
The above two vaccines are all bacteria-based vaccines, and their common disadvantage is that the antigen components injected into the body are many and complex, and many components are actually non-protective antigens
The existence of these non-protective antigen components interferes with the body's response to protective antigens (sometimes the response to non-protective antigens becomes the main response, while the protective antigen is a non-main response or even no response); The presence of antigens may bring about severe immune side effects, such as fever, immune diseases, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A preparation method and application of a non-toxic type 2 Streptococcus suis:

[0024] 1. Bacterial enrichment culture

[0025] Collect samples from the pharynx and nose of healthy pigs with cotton swabs, put into 0.5ml THB medium (Todd Hewitt Broth, Bacto TM ), after mixing on a vortex shaker, culture at 30°C for 6 hours.

[0026] 2. Purification and separation

[0027] The total DNA of the above-mentioned cultured bacteria was extracted by conventional methods as a template, and polymerase chain reaction (PCR) was carried out with primers designed for the type 2 capsular polysaccharide (cps2) gene, and the positive samples were determined to contain type 2 pig Streptococcus. A single colony was picked from the positive sample by streaking on a plate, and then cultured for enrichment. After PCR, Gram staining and serum agglutination tests, it was confirmed that the isolated strain was Streptococcus suis type 2, named 05JYS68.

[0028] 3. Animal experiments

[002...

Embodiment 2

[0033] A preparation method and application of a non-toxic type 2 Streptococcus suis:

[0034] 1. Bacterial enrichment culture

[0035] Collect samples from the pharynx and nose of healthy pigs with cotton swabs, put into 0.5ml THB medium (Todd Hewitt Broth, Bacto TM ), mixed on a vortex shaker, and incubated at 37°C for 4 hours.

[0036] 2. Purification and separation

[0037] The total DNA of the above-mentioned cultured bacteria was extracted by conventional methods as a template, and polymerase chain reaction (PCR) was carried out with primers designed for the type 2 capsular polysaccharide (cps2) gene, and the positive samples were determined to contain type 2 pig Streptococcus. A single colony was picked from the positive sample by streaking on a plate, and then cultured for enrichment. After PCR, Gram staining and serum agglutination tests, it was confirmed that the isolated strain was Streptococcus suis type 2, named 05JYS68.

[0038] 3. Animal experiments

[0039...

Embodiment 3

[0043] A preparation method of non-toxic type 2 Streptococcus suis:

[0044] 1. Bacterial enrichment culture

[0045] Collect samples from the pharynx and nose of healthy pigs with cotton swabs, put into THB culture medium (Todd Hewi tt Broth, Bacto TM ), mixed on a vortex shaker, and incubated at 39°C for 3 hours.

[0046] 2. Purification and separation

[0047] The total DNA of the above-mentioned cultured bacteria was extracted by conventional methods as a template, and polymerase chain reaction (PCR) was carried out with primers designed for the type 2 capsular polysaccharide (cps2) gene, and the positive samples were determined to contain type 2 pig Streptococcus. A single colony was picked from the positive sample by streaking on a plate, and then cultured for enrichment. After PCR, Gram staining and serum agglutination tests, it was confirmed that the isolated strain was Streptococcus suis type 2, named 05JYS68.

[0048] 3. Animal experiments

[0049] In order to c...

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PUM

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Abstract

The present invention relates to belongs to the field of bacteria, and is especially one avirulent type-II pig streptococcus strain and its preparation process and application in developing pig streptococcicosis vaccine. The avirulent type-II pig streptococcus strain is Streptococcus suis (Serotype 2)05JYS68 in the preservation number of CGMCC No. 1902. The preparation process includes the following steps: sampling from pharynx and nose of health pig, mixing with THB culture medium, static culture at 30-39 deg.c for 3-6 hr, extracting total DNA as template, PCR, plate streak, Gram staining and blood serum agglutination experiment to determine the strain.

Description

technical field [0001] The invention belongs to bacteria, specifically a strain of avirulent Streptococcus suis sero type 2 (Strep tococcus suis sero type 2) and its preparation method, and its application in the development of Streptococcus suis disease vaccine. Background technique [0002] Streptococcus suis type 2 (S. suis for short) is an important zoonotic pathogen. Pigs are generally susceptible to type 2 S. suis, and can cause acute sepsis, pneumonia, meningitis, arthritis, endocarditis, and acute death after infection. If not treated in time, the fatality rate can reach 40%. Pigs grow slowly and their storage time is greatly extended. In recent years, the prevalence of S. suis type 2 in pig herds in southern provinces of my country has become increasingly serious, and large-scale outbreaks have occurred from time to time, causing economic losses of one billion yuan each year. The pathogen infects humans and can lead to serious diseases such as meningitis and sepsi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/74A61K39/09A61P31/04C12R1/46A61K35/744
Inventor 唐家琪王长军郑峰潘秀珍
Owner 中国人民解放军南京军区军事医学研究所