Combined inactivated vaccine for avian influenza virus and fowl adenovirus

A dual inactivated vaccine, avian influenza virus technology, applied in the direction of vaccines, viruses, antiviral agents, etc., can solve the problems of increasing the manpower and material burden of chicken farms, affecting production performance, and increasing the susceptibility of chickens to diseases. , to achieve the effect of good safety, good immunogenicity and high titer

Active Publication Date: 2016-05-18
YEBIO BIOENG OF QINGDAO
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In recent years, H9 subtype avian influenza is a relatively common important disease that seriously threatens poultry; and chickens are more and more infected with I group 4 avian adenovirus, and this disease is easy to cause secondary infection disease, which is harmful to poultry. It brings a lot of troubles to commercial farmers, and the repeated use of vaccines, especially inactivated vaccines, not only increases the burden of manpower and material resources on the chicken f

Method used

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  • Combined inactivated vaccine for avian influenza virus and fowl adenovirus
  • Combined inactivated vaccine for avian influenza virus and fowl adenovirus
  • Combined inactivated vaccine for avian influenza virus and fowl adenovirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: the screening of YBAV-4 strain strain

[0022] 1. Epidemiological survey Since 2010, some broiler breeders, laying hens and Ma chickens in Shandong, Jiangsu and other regions have experienced a disease characterized by high mortality and anatomical manifestations of hepatomegaly and hydropericardium. The disease was initially diagnosed as hydropericardium hepatitis syndrome caused by group I group C-4 avian adenovirus through clinical investigation and laboratory testing. In 2010, the inventor successfully isolated a virus strain from the liver of a dead chicken with typical symptoms of inclusion body hepatitis and hydropericardium in a farm in Zibo, Shandong.

[0023] 2. Virus isolation Take the liver of sick and dead chicken and grind it, add sterilized saline at a ratio of 1:5 to make a suspension; after repeated freezing and thawing 3 times, centrifuge at 3000r / min for 30min, take the supernatant; add penicillin and Streptomycin at 10,000 IU / ml each, ...

Embodiment 2

[0032] Embodiment 2: the preparation of YBAV-4 strain virus seed

[0033] (1) Study on optimal culture conditions of chicken hepatocytes

[0034] 1. The effect of cell density on cell growth. Five different densities (10,000 to 50,000 / ml, 50,000 to 100,000 / ml, 100,000 to 150,000 / ml, 150,000 to 200,000 / ml, 20 to 25 10,000 / ml) chicken liver cells were inoculated into the same batch of 25cm 2 Cell bottle, 225cm 2 In cell flasks, 3000ml spinner bottles, and 10-layer cell factories, use the same batch of DMEM nutrient solution to culture under the same conditions, inoculate 5 bottles / 2 cells at each density, culture under the same conditions, and observe that the cells grow into a dense monolayer The time required and the shape of the cells.

[0035] 2, the influence of newborn bovine serum content on cell growth with the newborn bovine serum produced by the same manufacturer, add in the DMEM medium by the ratio of 6%, 8%, 10%, 12% respectively, cultivate the same batch of chick...

Embodiment 3

[0045] Example 3: Preparation of H9 subtype avian influenza, avian adenovirus (group I, type 4) antigen

[0046] 1. The use, maintenance conditions and methods of use of the Millipore ultrafiltration concentrator should be carried out according to the instructions provided by the manufacturer.

[0047] 2. Detection of concentration effect

[0048] 2.1 Antigen titer (HA and / EID) in the virus liquid before concentration was determined by retaining sample before concentration. 50 / TCID 50 ), during the concentration process, take samples at any time to measure the antigen titer (HA) of the concentrated solution, determine the concentration multiple, retain the sample of the concentrated solution after concentration, and measure the antigen titer (HA and / EID) in the concentrated solution 50 / TCID 50 ).

[0049] 2.2 Filtrate Antigen Detection Take the filtrate when the concentration is about to end (at this time, the concentration of the antigen in the concentrate is the highe...

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Abstract

The invention provides a combined inactivated vaccine for an avian influenza virus and a fowl adenovirus. An H9 subtype avian influenza virus QDY strain and an I-group 4-type aviadenovirus YBAV-4 new strain used by the vaccine are high in TCID50/EID50 valence and good in immunogenicity and can well withstand attacks of the H9 avian virus and various local separate viruses. The vaccine prepared by the invention is good in safety, and free of local and whole-body adverse effects caused by the vaccine. Through analysis of characters, a safety test and efficacy test data in a storage-life test, the combined vaccine is free of an obvious difference from a single vaccine of a similar product in effect, and is stable and effective; the efficacy test result proves that the antibodies of the combined vaccine and two single vaccines are kept at a high level and are faster in antibody generation of similar products; and a control group antibody is negative.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a dual inactivated vaccine of avian influenza virus and avian adenovirus. Background technique [0002] Avian influenza is caused by type A influenza virus and is mainly prevalent in animals and humans. It is an acute and highly contagious disease with high mortality. It occurs widely in all parts of the world and often mutates. It is harmful to humans and poultry. The industry is very dangerous. [0003] According to the epidemiological investigation of group I poultry adenovirus, the disease has a high incidence in chicken flocks in my country, can be transmitted through two ways, horizontal and vertical, and shows an increasing trend year by year. The host range of the disease is also becoming wider and wider. White-feathered broiler chickens, broiler breeders, laying hens, and yellow-feather chickens can all be infected. Especially after 20...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61K39/235A61P31/14A61P31/16C12N7/00C12N7/04C12R1/93
CPCA61K9/0019A61K9/107A61K39/12A61K2039/5252A61K2039/552A61K2039/70C12N7/00C12N2710/10034C12N2710/10261C12N2710/16021C12N2760/16021C12N2760/16061C12N2760/16134A61K2300/00
Inventor 李陆梅宫晓刘新文朱艳梅徐保娟
Owner YEBIO BIOENG OF QINGDAO
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