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Staphylococcus aureus vaccine and preparation method thereof

A staphylococcus, golden yellow technology, applied in the field of microbial vaccines, can solve problems such as narrow scope of application, poor immunogenicity, and no coverage of toxic proteins, and achieve the effect of reducing mortality and preserving immunogenicity

Inactive Publication Date: 2016-08-10
SICHUAN YUANDASHUYANG PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the immunogenicity of these methods is not as good as that of whole bacterial vaccines, most of the toxic proteins are not covered, the coverage of capsular polysaccharide is not enough, and the scope of application is narrow.

Method used

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  • Staphylococcus aureus vaccine and preparation method thereof
  • Staphylococcus aureus vaccine and preparation method thereof
  • Staphylococcus aureus vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] A preparation method of staphylococcus aureus vaccine, comprising the following steps:

[0038] (1) Inoculate the bacterial solution of Staphylococcus aureus standard strain ATCC 25923 on a blood plate, and incubate at 37°C for 24 hours;

[0039] (2) Pick 1 colony, inoculate it into high-pressure sterilized tryptone broth, shake and culture at 37°C for 18h to OD 600 0.6-0.8, the speed is 220rpm;

[0040] (3) Centrifuge at 3000rpm for 10min, discard the supernatant, resuspend the pellet with sterile PBS and centrifuge again, repeat 3 times to remove the medium and metabolites, and resuspend the pellet obtained after the last centrifugation with sterile PBS , so that its concentration is 1×10 9 / ml;

[0041] (4) Place the bacterial liquid on ice, and carry out radiation treatment with a gamma ray generating device, the radiation intensity is 10Gy / min, the radiation time is 10h, and the total dose is 6kGy;

[0042] (5) Aspirate 10 μl of the irradiated bacterial solution ...

Embodiment 2

[0046] Verification of immune protection of Staphylococcus aureus vaccine

[0047] Detect the protectiveness of Staphylococcus aureus vaccine in 4-6 week old Balb / c mice, each 10 of immunization group and control group, immunization group and control group are as shown in table 1:

[0048]

[0049]

[0050] Table 1

[0051] 2 weeks after the initial immunization, use the same dose in Table 1 for the second immunization injection; 3 weeks after the initial immunization, use the same dose in Table 1 for the last immunization injection; 2 weeks after the last immunization, intraperitoneally inject 0.5ml of lethal dose of Staphylococcus aureus ATCC 25923 to the mice , the number of bacteria is about 1×10 7 . The morbidity and death of the mice were observed, and the survival rates of the mice in the immunized group and the control group were counted after 2 weeks. Specific results such as figure 1 shown.

[0052] Depend on figure 1 It can be seen that the survival rate...

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Abstract

The invention discloses a staphylococcus aureus vaccine and a preparation method thereof. Staphylococcus aureus is inactivated through radiation and adjuvants are added, so that the proliferative activity of the staphylococcus aureus loses and the metabolic activity of the staphylococcus aureus is reserved; compared with thermal inactivation and inactivation with such chemicals as formaldehyde and the like, protein and polysaccharide antigens on the surface of the bacterium body (the staphylococcus aureus) can be well reserved, and immune response and immune protection can be comprehensively and obviously induced. The preparation method is simple and convenient and is suitable for the large-scale preparation of the staphylococcus aureus vaccine; the preparation method is not only suitable for preparing the staphylococcus aureus vaccine, but also capable of serving as a rapid response strategy of bio-threat or newly emerging pathogens; especially, for pathogens which are not clear in immune response mechanism, a candidate vaccine can be rapidly prepared by virtue of the method; and the method can shorten research and development time and can reduce research and development cost.

Description

technical field [0001] The invention relates to the field of microbial vaccines, in particular to a Staphylococcus aureus vaccine and a preparation method thereof. Background technique [0002] Staphylococcus aureus is an important pathogen of humans, causing many serious infections. Staphylococcus aureus generally exists in food and can cause food poisoning in humans through transmission. It can also cause pneumonia, pseudomembranous enteritis, pericarditis, etc., and even systemic infections such as sepsis and sepsis, which are seriously harmful. For the preventive measures of Staphylococcus aureus, the main measures are to strengthen infection control, use antibiotics, remove the carried Staphylococcus aureus and immunological methods. Among them, the immunological method is mainly to inoculate Staphylococcus aureus vaccine to improve the antibody resistance in the body and enhance the performance of cellular immunity, which is a very effective method. [0003] The Chin...

Claims

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Application Information

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IPC IPC(8): A61K39/085A61K41/00A61P31/04
CPCA61K39/085A61K41/10A61K2039/55505
Inventor 沈玉保李竹石魏宪义蒋德席
Owner SICHUAN YUANDASHUYANG PHARM CO LTD
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