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A porcine epidemic diarrhea vaccine strain and vaccine containing same

A technology for swine epidemic diarrhea and vaccine virus, which is applied in the field of virus strains and vaccines containing the virus strains, can solve the problems of difficulty in meeting the requirements of neutralizing antibody levels, poor stability of vaccine virus strains, and insufficient immunogenicity, and achieves prevention of pigs. Epidemic diarrhea infection, good immunogenicity, good passive protection effect

Active Publication Date: 2020-06-09
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the poor stability of the vaccine virus strain, difficulty in cultivating, and insufficient immunogenicity, it is difficult to meet the requirements of neutralizing antibody levels after animal vaccination, resulting in frequent outbreaks of porcine epidemic diarrhea in immunized pig farms

Method used

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  • A porcine epidemic diarrhea vaccine strain and vaccine containing same
  • A porcine epidemic diarrhea vaccine strain and vaccine containing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The preparation of embodiment 1 porcine epidemic diarrhea vaccine strain

[0023] (1), Cloning Screening of Porcine Epidemic Diarrhea Vaccine Strain ZM-PE56

[0024] Porcine epidemic diarrhea vaccine strain ZM-PE56 was obtained by 5 plaque clones and 6 passages. Its cloning method is as follows:

[0025] Form a monolayer of Vero cells and plate a 6-well plate.

[0026] Take 100 μL of PEDV virus liquid, dilute it with diluent according to 10-fold serial dilution, and dilute 10 -6 、10 -5 、10 -4 、10 -3 、10 -2 The diluted virus solution was inoculated in a 6-well plate covered with a monolayer of Vero cells, each well was connected with 0.5ml, and a negative control well without the virus was set at the same time.

[0027] Discard the cell culture medium in the cell culture plate before inoculation, rinse the six-well plate twice with plate washing solution, and then discard it.

[0028] After adsorption at 37°C for 1.5h (shake once every 30 minutes), the virus liqu...

Embodiment 2

[0042] The preparation of embodiment 2 porcine epidemic diarrhea vaccine

[0043](1) Antigen preparation

[0044] PEDV production seeds (virus content ≥ 10 6.0 TCID 50 / ml), according to 0.1% ~ 1% of the culture volume inoculated to form a single layer of Vero cells, add DMEM maintenance solution for culture, when 75% of the cells have lesions (about 72h), freeze and thaw twice to harvest the virus liquid, centrifuge Or the supernatant collected after filtration is the antigen. Harvested antigens were serially diluted 10-fold and inoculated into 96-well plate Vero cell monolayers, each dilution was inoculated into 8 wells, observed for 4 days, and the virus content was calculated according to the Reed-Muench method formula. After determination, the content of one batch of antigenic virus cultivated was 10 7.5 TCID 50 / ml.

[0045] (2) The preparation of the finished vaccine product uses pure water as a solvent, and prepares hydrolyzed gelatin containing 3%, 1% casein hyd...

Embodiment 3

[0048] Embodiment 3 stability test

[0049] This embodiment is used to illustrate the stability of the prepared vaccine of the present invention, and specific operations are as follows:

[0050] The two batches of vaccines 14001 and 14002-CG prepared in Example 2 were placed at 37°C for 7 days to measure the change in toxicity, and the results showed that the toxicity of the former decreased by 10 0.7 TCID 50 , while the latter drops by 10 1.5 TCID 50 , indicating that the addition of suitable concentrations of hydrolyzed casein, arginine hydrochloride, sodium glutamate and other ingredients in the new protective agent can significantly improve the stability of the vaccine.

[0051] The two batches of vaccines 14001 and 14002-CG prepared in Example 2 were placed in a refrigerator at -15°C, and the changes in toxicity were measured at 0, 12, 24 and 36 months respectively. As a result, at different time points, the virus content of each vaccine in 14001 batches of vaccines w...

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Abstract

The invention relates to an animal vaccine, and particularly discloses a porcine epidemic diarrhea vaccine strain and a vaccine containing the porcine epidemic diarrhea vaccine strain. The porcine epidemic diarrhea vaccine strain adopts porcine epidemic diarrhea virus attenuated by separation and purification as virus seed, and is freeze-dried by suitable protecting agent; the virus is injected topigs by muscle injection, thus high neutralizing antibody level is generated and passive protection is provided for suckling pigs; the vaccine has the advantages of simple preparation technique, stable property, and good immunogenicity; besides, the vaccine can effectively prevent porcine epidemic diarrhea virus infection.

Description

technical field [0001] The invention relates to animal vaccines, in particular to a strain of porcine epidemic diarrhea and a vaccine containing the strain. Background technique [0002] Porcine epidemic diarrhea (PED) is an acute gastrointestinal infectious disease of pigs caused by porcine epidemic diarrhea virus (PEDV), characterized clinically by vomiting, diarrhea, and dehydration. During the autumn and winter of 2010, PED broke out in many provinces in China, and soon spread to a national pandemic. During this period, the incidence rate of suckling piglets and nursery pigs was 80% to 100%, and the mortality rate was as high as 50% to 90%. Prevention of the disease mainly relies on vaccination. [0003] Porcine epidemic diarrhea vaccines sold in the market are mainly porcine epidemic diarrhea and porcine transmissible gastroenteritis dual live vaccines prepared by traditional techniques. Due to the poor stability of vaccine virus strains, difficulty in cultivating, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/215A61P31/14
CPCA61K39/12C12N7/00C12N2770/20021C12N2770/20034
Inventor 马良潘文许磊李晓冉邵葳周建民
Owner CHINA ANIMAL HUSBANDRY IND
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