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Preparation method and application of Newcastle disease virus infected immune complex vaccines

An immune complex and Newcastle disease virus technology, applied in the field of preparation and application of immune complex vaccines for Newcastle disease virus infection, to achieve the effect of inhibiting Newcastle disease virus infection

Inactive Publication Date: 2011-11-09
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although now widely vaccinated against the disease, it still causes heavy economic losses in the poultry industry from time to time and is currently one of the most important and dangerous poultry diseases
At present, the main measure to control the disease is through vaccination, but both inactivated vaccines and live vaccines have their own advantages and disadvantages. Facing the shortcomings of conventional Newcastle disease vaccines, many researchers are committed to the development of new vaccines

Method used

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  • Preparation method and application of Newcastle disease virus infected immune complex vaccines
  • Preparation method and application of Newcastle disease virus infected immune complex vaccines
  • Preparation method and application of Newcastle disease virus infected immune complex vaccines

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Preparation of antigens and inactivated vaccines

[0019] The Shanghai strain of Newcastle disease virus was used as the seed virus, and it was inoculated through the allantoic cavity. The specific method was: take 9-10-day-old SPF chicken embryos (purchased from Beijing SPF chicken farm), and draw the air cells and embryos with an egg inspection lamp. Once positioned, the egg is placed transversely on the cuspid with the embryo positioned downward. Sterilize with iodine tincture and alcohol at 1 / 2 of the long diameter of the egg. Make a small hole with a steel cone, pierce the needle about 1.5 cm, inject 0.1-0.5 mL of virus solution, seal the hole with molten paraffin, and place it in an incubator to continue incubation. Take out the eggs at 24h, discard the dead chick embryos, put the rest into the incubator, and lighten the eggs again within 48-60h, put the live chick embryos in a 4°C refrigerator, take out the allantoic fluid after 24h, and use Hemagglutination te...

Embodiment 2

[0022] Preparation of egg yolk antibody IgY

[0023] After immunizing SPF laying hens four times with the inactivated vaccine prepared in Example 1, the laid eggs were collected and precipitated with PEG and ammonium sulfate to prepare egg yolk antibody (IgY). The specific process is as follows:

[0024] Carefully separate the yolk from the egg, mix 10ml of egg yolk solution and 40ml of 0.1mol / L phosphate buffered saline (PBS) at pH 7.4, centrifuge at 4500r / min for 30min, and add PEG-8000 to the supernatant to make the concentration of 6wt%, stirred at room temperature to dissolve, centrifuged at 12000r / min at 4°C for 15 minutes, the obtained egg yolk antibody IgY was dissolved in 10mL of PBS, and then PEG-8000 was slowly added to make the concentration 15wt%, and the IgY was purified by centrifugation after precipitation. Dissolve in 50 mL of PBS, slowly add saturated ammonium sulfate to reach a final concentration of 50% (v / v), centrifuge at 12,000 r / min at 4°C for 15 minut...

Embodiment 3

[0026] Preparation of immune complex vaccine

[0027] The inactivated vaccine prepared in Example 1 and the antibody prepared in Example 2 were mixed in equal volumes and incubated at 20° C. for 1 hour to obtain the immune complex vaccine of the present invention.

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Abstract

The invention discloses a preparation method and application of Newcastle disease virus infected immune complex vaccines. The preparation method comprises the following steps of: (1) preparing antigens and inactivated vaccines by using Newcastle disease virus Shanghai strains as seed viruses; (2) transferring antibodies in chicken blood serum to yolk to form yolk antibodies IgY; and (3) mixing the prepared inactivated vaccines and the yolk antibodies IgY in equal volume, and hatching to obtain the immune complex vaccines. By combining passive immune (vaccine) and active immune (specific antibody) measures, animals are protected from the infection of pathogenic microbes in time for a long time.

Description

technical field [0001] The invention relates to the preparation and application of DNA vaccines, in particular to a preparation method and application of a Newcastle disease virus infection immune complex vaccine. Background technique [0002] Newcastle disease (ND), also known as Asian chicken plague, pseudo-fowl plague, etc., is an acute and highly contagious infectious disease caused by Newcastle disease virus (NDV). The typical characteristics of Newcastle disease are increased body temperature, loss of appetite, drooping neck or drooping wings, sick chickens cough, difficulty breathing, and the crop is full of liquid contents, often with sour and smelly liquid flowing out of the mouth, and some sick chickens also appear. Neurological symptoms, diarrhea, cloacal hemorrhage, necrosis, glandular gastric papilla, glandular stomach and gizzard junction and duodenal bleeding, chronic cases often have respiratory symptoms or neurological symptoms. Although now widely vaccinat...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P31/14A61K39/17
Inventor 易建中刘成倩
Owner SHANGHAI ACAD OF AGRI SCI
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