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Novel cairna moschata adenovirus strain, novel cairna moschata adenovirus inactivated vaccine, and preparation method for inactivated vaccine

A virus inactivation, Muscovy duck gland technology, applied in biochemical equipment and methods, viruses, antiviral agents, etc., can solve problems such as economic losses in the Muscovy duck breeding industry, achieve good commercial development prospects, and reduce economic losses. Effect

Active Publication Date: 2019-12-13
SHANDONG SINDER TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After the isolation and identification of the pathogen and the artificial infection test of the virus, it was preliminarily shown that the pathogen of the disease was a new type of Muscovy duck adenovirus different from DAdV-2. Some experts named the virus Duck adenovirus type 3 (DAdV -3), but this nomenclature has not been accepted by the International Committee on Taxonomy of Viruses (ICTV)
Through the epidemiological investigation of the new Muscovy duck adenovirus, it was found that the disease is prevalent in almost all Muscovy duck breeding areas in my country, causing great economic losses to the Muscovy duck breeding industry
At present, there is no commercial vaccine (or antibody) for effective prevention and treatment of the disease at home and abroad, and it is imminent to develop a safe and effective vaccine and antibody

Method used

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  • Novel cairna moschata adenovirus strain, novel cairna moschata adenovirus inactivated vaccine, and preparation method for inactivated vaccine
  • Novel cairna moschata adenovirus strain, novel cairna moschata adenovirus inactivated vaccine, and preparation method for inactivated vaccine
  • Novel cairna moschata adenovirus strain, novel cairna moschata adenovirus inactivated vaccine, and preparation method for inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Isolation and identification of Muscovy duck novel adenovirus GD MM strain

[0037] 1 Epidemiological survey

[0038]Since 2014, young muscovy ducks in Guangdong, Fujian, Anhui and other regions have developed infectious diseases with symptoms such as liver swelling and congestion, varying degrees of hemorrhage or necrosis on the surface, brittle texture, and liver color turning yellowish or white. . The disease usually affects Muscovy ducks under 1 month old, and the morbidity and mortality show a certain age-related correlation. At present, the prevalence of the disease is found in almost all Muscovy duck breeding areas. After clinical investigation and laboratory testing, the preliminary diagnosis was a new type of duck adenovirus. In 2018, a strain of the virus was successfully isolated from a group of 20-day-old Muscovy ducks in a Muscovy duck farm in Maoming, Guangdong.

[0039] 2 Virus isolation:

[0040] Select 50-100g of dead muscovy duck liver,...

Embodiment 2

[0064] Embodiment 2: the preparation of Muscovy duck novel adenovirus GD MM strain seed

[0065] Take the third-generation embryotoxic allantoic fluid of Muscovy duck new adenovirus, filter it through a 0.22 μL filter, inoculate LMH cells growing to a single layer at a ratio of 3-5% of the culture medium, and discard the original culture medium, and adsorb for 1 hour at 37°C Discard the virus liquid, and then add bovine serum-free DMEM / F12 virus maintenance solution, 37 ° C, 5% CO 2 Under the condition of static culture for 72-96h, the virus fluid was harvested. The LMH cells were subcultured to proliferate the F5-F9 generation until more than 80% of the cells became lesioned, and the lesion time was controlled within 72-96 hours to harvest the virus liquid. Measure the virus content of this strain 5-9 generations, the results are shown in Figure 7 . Mix the sterile virus liquid tested in the same generation, quantitatively aliquot it, and store it in a -80°C refrigerator....

Embodiment 3

[0066] Example 3: Preparation of muscovy duck novel adenovirus GD MM strain antigen

[0067] LMH cell culture: Take LMH cells grown to a dense monolayer, discard the original culture medium, wash the cells with sterile PBS, digest with 0.25% trypsin, discard the trypsin solution, add an appropriate amount of 8% fetal bovine serum The DMEM / F12 cell culture medium was used to mix the cells, and the cells were passaged at a ratio of 1:4-1:5, at 37°C, 5% CO 2 cultured to a monolayer of cells.

[0068] Inoculation: Discard the original culture medium of the above-mentioned LMH cells covered with a single layer, inoculate LMH cells at a final concentration of 1‰-1%, and place at 37°C in 5% CO 2 The incubator under the condition continued to cultivate for 72-96h.

[0069] Toxic collection: After receiving the poison, observe twice a day, and record the cell lesions. Harvest the virus liquid when the cytopathy reaches more than 80%, and store it at -20°C.

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Abstract

The invention disclose a novel cairna moschata adenovirus strain, a novel cairna moschata adenovirus inactivated vaccine, and a preparation method for the inactivated vaccine. The adenovirus strain iscollected in the China Center for Type Culture Collection in the Wuhan University on the July 30th, 2019 with a collection number of CCTCC No:V201952. The invention establishes a preparation method for a corresponding inactivated vaccine on the basis of the novel cairna moschata adenovirus. The inactivated vaccine prepared with the method can effectively prevent and treat a novel cairna moschataadenovirus disease, has a good commercial development prospect, solves the problem that no effective solutions can be provided when the novel cairna moschata adenovirus begins to spread on a large scale at present, and greatly reduces the economic loss of cairna moschata breeding industry due to the novel cairna moschata adenovirus disease.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a novel muscovy duck adenovirus strain, an inactivated vaccine and a preparation method thereof. Background technique [0002] Avian adenovirus is one of the common infectious disease pathogens in poultry. Most avian adenoviruses can exist and replicate in healthy poultry without clinical symptoms or very mild symptoms, but adenoviruses can become conditional pathogens during mixed infection; sometimes avian adenoviruses can also be used as primary pathogens and can cause Clinical and pathological syndromes of chickens, ducks, geese and other poultry or wild birds, including hydropericardium, hepatitis, aplastic anemia, hemorrhage, mild respiratory diseases and other diseases of poultry. [0003] Duck (Anas platyrhynchos domestica)-derived adenoviruses include duck-derived adenovirus type A (DAdV-A) of the AT-rich adenovirus genus and duck-derived adenovir...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/235A61P31/20C12R1/93
CPCA61K39/12A61K2039/5252A61P31/20C12N7/00C12N2710/10221C12N2710/10234C12N2710/10251C12N2710/10263
Inventor 罗济冠常娓娓王学波刘宁郭春丽李朝阳
Owner SHANDONG SINDER TECH
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