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Newcastle disease diad vaccine and preparation method thereof

A doublet vaccine, chicken Newcastle disease technology, applied in the field of animal vaccine preparation, can solve problems such as vaccines that have not yet seen chicken Newcastle disease disease, achieve the effects of preventing and treating virus infection, improving immunity, and overcoming the prevention of a disease

Inactive Publication Date: 2014-01-08
河南亚卫动物药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the prior art, there have been many studies on the use of HN protein alone to prepare subunit vaccines for the prevention of chicken Newcastle disease, and there are many reports on the use of chicken α-interferon (α-IFN) as a vaccine to improve chicken immunity or prevent and treat diseases, but not yet. See related reports on the combination of the two to jointly prepare a vaccine to prevent Newcastle disease

Method used

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  • Newcastle disease diad vaccine and preparation method thereof
  • Newcastle disease diad vaccine and preparation method thereof
  • Newcastle disease diad vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] This example focuses on the preparation process of chicken Newcastle disease doublet vaccine.

[0035] The preparation process of the doublet vaccine of the present invention generally comprises the following steps:

[0036] (1) Using PCR and RT-PCR techniques to clone the chicken α-IFN gene and the chicken Newcastle disease virus HN gene respectively;

[0037] (2) Cloning the HN gene and the chicken α-IFN gene into the eukaryotic expression vector pcDNA3.1, constructing the recombinant eukaryotic double expression vector pcDNA3.1-IFN-HN, screening positive positive clones by PCR and enzyme digestion;

[0038] (3) BHK-21 cells were transfected with the recombinant plasmid pcDNA3.1-IFN-HN, and then ZeocinTM resistance screening was performed;

[0039] (4) Perform indirect immunofluorescence (IFA) on the screened positive cells and use Western-blotting technology to detect the expression of the target protein HN and α-IFN in BHK-21 cells;

[0040] (5) Use protein purifi...

Embodiment 2

[0102] In order to further test the preventive effect of the doublet vaccine prepared in the present invention against Newcastle disease virus, the inventors conducted further animal experiments.

[0103] Select 80 5-day-old healthy chicks with ND-negative serology, and randomly divide them into 4 groups, namely: blank control group (0.5ml / bird with conventional bacterial vaccine), low-dose doublet vaccine group (0.3ml / ), middle-dose group (0.5ml / rat), high-dose group (0.8ml / rat), 20 rats in each group.

[0104] Utilize the method for preparing chicken Newcastle disease HN and α-IFN dyad vaccine provided in Example 1, prepare dyad vaccine, and inoculate it subcutaneously in the neck of chicks, and the specific comparative treatment is shown in the following table

[0105] Table 1 Grouping and treatment of experimental chickens

[0106] group Dose / ( ml / piece) Number of test chickens / piece Route of administration Blank control group 0.5 20 Inoculatio...

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Abstract

The invention belongs to the technical field of animal vaccine preparation, and particularly relates to a diad vaccine regarding HN protein and chicken alpha interferon as effective components, and a preparation method thereof. The diad vaccine is prepared by transfection of BHK-21 cells and fully expressing obtained target protein by using HN genes and chicken alpha interferon genes through the utilization of a genetic engineering technology, and mixing the target protein with an adjuvant. When the diad vaccine prepared through the utilization of the expression proteins of the HN genes and the chicken alpha interferon genes is injected in a chicken body, the HN proteins and the alpha interferon proteins play respective biological roles in the chicken body, so that the effects of positively preventing and treating other virus infections are achieved while the virus infection of the newcastle disease is prevented. The vaccine is totally safe and reliable, is capable of inducing an organism to generate specific antibodies continuously for a long time and improving gallinaceous immunity, and well overcomes the disadvantage that the traditional vaccine only can prevent one disease.

Description

technical field [0001] The invention belongs to the technical field of animal vaccine preparation, and in particular relates to a doublet vaccine with HN protein and chicken α-interferon (α-IFN) as active ingredients and a preparation method thereof. Background technique [0002] In the process of poultry breeding, with the continuous emergence of new strains and mutant strains of pathogenic bacteria, it has gradually become difficult for drugs that have good control effects on certain livestock and poultry diseases to achieve satisfactory therapeutic effects. In recent years, with the increasingly serious harm of viral diseases in the process of poultry farming, my country has invested huge manpower, material and financial resources in the preparation of vaccine products for disease prevention and control, which has played a key role in controlling the spread of large-scale epidemics of diseases. effect. [0003] Newcastle disease (ND) is a highly contagious, acute septic ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/17A61P31/14A61K38/21
Inventor 郭芳茹杨慧敏张遂平杨果王菊萍董战杰王黎明
Owner 河南亚卫动物药业有限公司
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