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Duck virus hepatitis strains and inactivated vaccine

A technology for duck viral hepatitis and inactivated vaccines, which is applied in antiviral agents, viruses/bacteriophages, biochemical equipment and methods, etc., can solve the problems of breeding losses, local outbreaks of duck viral hepatitis, etc., and achieve good immune effects , normal feeding, good safety effect

Active Publication Date: 2011-06-08
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immune ducklings with attenuated vaccines, affected by the immune function of ducklings and maternal antibodies, caused local outbreaks of duck viral hepatitis, causing huge losses to the breeding industry

Method used

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  • Duck virus hepatitis strains and inactivated vaccine
  • Duck virus hepatitis strains and inactivated vaccine
  • Duck virus hepatitis strains and inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Preparation of Duck Viral Hepatitis Virus Strain (DHV YC)

[0038](1) Disease material treatment: Duck farms in Henan where duck viral hepatitis occurred, collected duck liver disease materials with obvious lesions, washed 3 times with sterilized PBS containing 100IU / ml penicillin and 100mg / ml streptomycin double antibodies , Dilute and grind with sterilized PBS 5 times the weight of the disease material. Freeze and thaw the prepared homogenate at -20°C, freeze and thaw twice at 20°C, centrifuge at speeds of 3000r / min and 8000r / min for 15min, and shake the supernatant with chloroform at room temperature for 10-15min. The supernatant was collected by centrifugation, filtered through a 0.22 μm filter membrane, aliquoted, and stored at -70°C.

[0039] (2) Establishment of chicken embryo-adapted strain of duck hepatitis virus: the virus suspension obtained in step (1) is diluted 1:100 with sterilized PBS, and 20 pieces of 9-day-old SPF chicken embryos are inoc...

Embodiment 2

[0040] Example 2 Preparation of Inactivated Vaccine of Duck Viral Hepatitis

[0041] (1) Dilute the DHV YC strain 100 times with sterilized PBS, inoculate it into 9-day-old SPF chicken embryos, and collect the chicken embryo fluid that died within 48-96 hours and had obvious lesions.

[0042] (2) Inactivation: add 0.05% (mass percentage) of β-propiolactone to the qualified chicken embryo liquid, inactivate at 4°C for 24h, and then hydrolyze it for 2h at 37°C in a water bath, as an antigen for making seedlings liquid.

[0043] (3) Preparation of vaccine by emulsification: oil phase preparation: take 94ml of white oil, 6ml of Siben-80, mix well and add 1.5g of aluminum stearate. Preparation of aqueous phase: Take 20ml of antigen solution, 3.6ml of Tween-80, 1.5g of immune enhancer isoprinosine, 4.9ml of sterilized saline, and mix well. Take 100ml of the oil phase and place it in an emulsification tank, pre-emulsify at 7500r / min for 3min, and slowly add 50ml of water phase a...

Embodiment 3

[0045] Example 3 Test of Duck Viral Hepatitis Vaccine

[0046] (1) Physical properties

[0047] Appearance milky white emulsion

[0048] Dosage Form Water-in-Oil

[0049] Viscosity Use a 1ml straw (outlet diameter: 1.2mm) to suck up the vaccine at about 25°C, let it flow out vertically and naturally, record the time required for the outflow of 0.4ml, which is 4.8s, which meets the regulations.

[0050] Stability Store at 37°C for 21 days or put 5ml of the vaccine into a centrifuge tube and centrifuge at 3000r / min for 15min without breaking the emulsion.

[0051] (2) Sterility test was carried out according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and there was no bacterial growth.

[0052] (3) Safety inspection Take 10 1-day-old Cherry Valley ducks, inject 1ml of vaccine subcutaneously in each neck, observe for 14 days, no local and systemic adverse reactions caused by vaccine injection appear.

[0053] (4) Efficacy test 10 1-day-old Cherry Val...

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Abstract

The invention discloses methods for preparing a duck virus hepatitis strain and an inactivated vaccine. The strain is obtained with the method comprising the following steps of: collecting duck hepatitis material with obvious pathological changes in a duck farm where duck virus hepatitis prevails, washing with bi-anti-sterilization PBS, diluting and grinding, freeze thawing twice and performing centrifugal separation; diluting virus separated by sterilization PBS, vaccinating chick embryo, selecting chick embryo with classic pathological changes that is dead 48-96h after vaccination, respectively gaining chick embryo liquid, transferring for 18-25 generations continuously to obtain a duck virus hepatitis virus chick embryo suitable strain. The strain can be in production, vaccinated in the chick embryo to gain chick embryo liquid, and is emulsified after vaccination to prepare the inactivated vaccine. The inactivated vaccine is high in safety, has good immune efficacy to young ducks, long immune time for breeding ducks and high level of antibody of breeding ducks, and the young ducks bred by the breeding ducks can resist duck virus hepatitis strains.

Description

technical field [0001] The invention relates to a preparation method of a duck viral hepatitis virus and an inactivated vaccine, more specifically, the invention relates to a duck viral hepatitis virus and an inactivated vaccine, and belongs to the technical field of biological products. Background technique [0002] Duck virus hepatitis (DVH) is an acute fatal infectious disease of ducklings. The disease has an acute onset and rapid spread, mainly causing ducklings, with a morbidity and mortality rate as high as 100%. The disease first occurred in the United States, and then became popular in the United Kingdom, Canada, Germany, Italy, India and other countries. Since duck hepatitis virus (DHV) was first isolated in Shanghai in 1958 in my country, it has been prevalent in many provinces and cities across the country, causing huge economic losses, and has become an important threat to the healthy development of the duck industry. [0003] Virus particles of duck viral hepa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/02A61K39/29A61P1/16A61P31/14C12R1/93
Inventor 张许科孙进忠乔荣岑
Owner PU LIKE BIO ENG
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