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

A technology of egg drop syndrome and vaccine composition, which is applied in the field of vaccine composition for the prevention of poultry egg drop syndrome, can solve problems such as no poultry egg drop syndrome subunit vaccines are on the market, and achieve good immunogenicity and biological good safety effect

Active Publication Date: 2018-10-16
PU LIKE BIO ENG
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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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Effect test

Embodiment approach

[0048] 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 Antigens, avian adenovirus antigens, avian reovirus antigens, Escherichia coli antigens, Avian bacillus paragallinarum antigens, Mycoplasma synoviae antigens, Mycoplasma gallisepticum antigens, Pasteurella multocida antigens, Marek's virus antigens, avian brain Myelitis virus antigen, chicken infectious laryngotracheitis virus antigen.

[0049] As an embodiment of the present invention, the vaccine composition also includes one or more of the group consisting of the following antigens: chicken Newcastle disease virus inactivated antigen, avian influenza virus inactivated antigen, chicken infectious bronchitis virus inactivated antigen Live antigen, chicken infectious bursal disease virus su...

Embodiment 1

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

[0062] 1. Extraction of EDSV virus DNA

[0063] The plasmid extraction kit was purchased from Tiangen Biotech; T4 DNA 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 ...

Embodiment 2

[0072] Preparation of embodiment 2 Penton protein

[0073] Inoculate the pET28a-EDSV-Penton / 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 OD 600 = 0.4 to 0.6, stand 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 rel...

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Abstract

The invention relates to a vaccine composition. The vaccine composition comprises an immunological dose of egg drop syndrome virus Penton protein or an immunological dose of a live vector containing arecombinant egg drop syndrome virus Penton 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 belongs to the field of biomedicine, and in particular relates to a vaccine composition for preventing poultry egg drop syndrome, a preparation method and application. Background technique [0002] Egg drop syndrome (EDS) is an infectious disease caused by Egg drop syndrome virus (Egg drop syndrome virus, EDSV), characterized by decreased egg production and deformed eggs. During the peak period of laying eggs, huge economic losses have been brought to laying poultry farming. [0003] EDSV belongs to group III of avian adenoviruses, has typical adenovirus morphology, has no capsule, has erythrocyte agglutinating activity, and proliferates in avian oviducts. The virus genome is a linear double-stranded DNA of about 33kb, with a single serotype, and there is no significant difference in antigenicity between strains. Virus particles are composed of structural proteins. The diameter of the nucleocapsid is 70-80nm, which is icosahedral and sy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/235A61K39/39A61P31/20
CPCA61K39/12A61K39/39A61K2039/552C12N2710/10234
Inventor 田克恭张盼涛逄文强孙进忠张许科
Owner PU LIKE BIO ENG
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