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Vaccine composition for preventing egg drop syndrome, and preparation method and application of vaccine composition

一种减蛋综合征、疫苗组合物的技术，应用在预防禽减蛋综合征的疫苗组合物领域，能够解决未有禽减蛋综合征亚单位疫苗上市等问题，达到免疫原性好、生物安全性好的效果

Active Publication Date: 2018-10-16
PULIKE BIOLOGICAL ENG INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no subunit vaccine for poultry egg drop syndrome has been marketed

Method used

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  • Vaccine composition for preventing egg drop syndrome, and preparation method and application of vaccine composition
  • Vaccine composition for preventing egg drop syndrome, and preparation method and application of vaccine composition
  • Vaccine composition for preventing egg drop syndrome, and preparation method and application of vaccine composition

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Experimental program
Comparison scheme
Effect test

Embodiment approach

[0048] The term "combined vaccine" refers to a vaccine prepared from a virus mixture of the avian egg drop syndrome virus of the present invention and at least one different virus. The term "composite vaccine" refers to a vaccine prepared from the avian egg drop syndrome virus and bacteria of the present invention. For example, the avian egg drop syndrome virus of the present invention can be combined with chicken Newcastle disease virus, chicken infectious bronchitis virus, avian influenza virus, chicken infectious bursal disease virus, avian adenovirus, avian reovirus and / or Escherichia coli , Avian bacillus paragallinarum, Mycoplasma synovialis, Mycoplasma gallisepticum mixed or combined. As an embodiment of the present invention, the vaccine composition also includes one or more of the following antigens: chicken Newcastle disease virus antigen, avian influenza virus antigen, chicken infectious bronchitis virus antigen, chicken infectious bursal disease virus antigen Anti...

Embodiment 1

[0061] Example 1 Construction of pET28a-EDSV-Fiber expression vector

[0062] 1. Extraction of EDSV virus DNA

[0063] The plasmid extraction kit was purchased from Tiangen Biotech; T4DNA Ligase was purchased from BioLab; the pET28a plasmid was purchased from Novagen; the agarose gel recovery kit was purchased from Tianze Biotech, and other reagents were of analytical grade.

[0064] According to the instructions of the virus DNA extraction kit, take 0.2ml of poultry egg drop syndrome virus solution into a sterile 1.5ml centrifuge tube, add 0.4ml of VB to the sample solution, vortex and mix, and let stand at room temperature for 10 minutes. Add 0.45ml AD buffer to the sample solution and mix vigorously. Put the VB column into a 2ml collection tube, add 0.6ml of the mixed solution to the VB column, centrifuge at 14000g for 1 minute, add the remaining mixed solution to the VB column, and centrifuge at 14000g for 1 minute, discard the 2ml collection tube, put the VB column in P...

Embodiment 2

[0072] The preparation of embodiment 2 Fiber protein

[0073] Inoculate the pET28a-EDSV-Fiber / pG-Tf2 / E.Coli BL21 (DE3) strain containing prepared in Example 1 into the LB medium containing 50-100 μg / ml kanamycin and 20 μg / ml chloramphenicol , while the LB medium contains 5-10 ng / ml tetracycline for inducing the expression of molecular chaperones, the inoculum size is 1% (V / V), and the culture is shaken at 37°C. When OD600=0.4~0.6, place it at 28°C for 30 minutes. Isopropyl-β-D-thiogalactopyranoside (IPTG) was added to make the final concentration 0.1-1.0 mM, and cultured with shaking at 28°C for 24 hours. After the cultivation, the cells were collected and resuspended with PBS (sodium chloride, 8g, potassium chloride, 0.2g, disodium hydrogen phosphate, 1.44g, potassium dihydrogen phosphate, 0.24g, adjusted to pH 7.4, constant volume 1L) Bacteria were sonicated and centrifuged to obtain the supernatant. The content of soluble target protein in the expression product is relat...

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Abstract

The invention relates to a vaccine composition. The vaccine composition comprises an immunological dose of egg drop syndrome virus Fiber protein or an immunological dose of a live vector containing arecombinant egg drop syndrome virus Fiber protein gene, and a veterinary acceptable carrier. The vaccine composition can effectively protect the occurrence of egg laying of chickens, and can completely prevent against wild strains of egg drop syndrome virus in different regions.

Description

technical field [0001] The invention relates to a vaccine composition for preventing poultry egg drop syndrome, a preparation method and application thereof, and belongs to the field of biomedicine. Background technique [0002] Egg Drop Syndrome Virus (EDSV) belongs to avian adenovirus group III. After the virus infects poultry, there are no obvious clinical symptoms, or mild diarrhea can be seen. It often occurs in poultry during the peak egg production period. Produce soft-shell, thin-shell, and shellless eggs, and the egg production rate drops severely, causing serious economic losses. [0003] EDSV has a typical shape of adenovirus, without capsule, has hemagglutinating activity, and proliferates in the oviduct of avian. The virus genome is a linear double-stranded DNA of about 33kb. The virus particles are composed of structural proteins. The diameter of the nucleocapsid is 70-80nm, which is icosahedral and symmetrical, and the DNA is enclosed in the capsid. The nucl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/235A61P31/20
CPCA61K39/12C12N2710/10234A61K2039/552A61P31/20A61K2039/5252C12N2710/10034C12N2720/10034C12N2760/16034C12N2760/18134C12N2770/20034A61K2039/55566A61K2039/70A61K39/235A61K39/17A61K39/145A61K39/215
Inventor 田克恭程艺逄文强孙进忠张许科
Owner PULIKE BIOLOGICAL ENG INC
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