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Raccoon dog parvoviral enteritis and canine distemper combined inactivated vaccine and preparation method thereof

A dual inactivated vaccine and parvovirus technology, which is applied in the field of veterinary biological products, can solve the problems of poor immunogenicity of inactivated vaccines, increase the workload of farmers, and the severe stress response of raccoon dogs, and achieve the automation of cultivation methods , Small training area and no need for production personnel

Active Publication Date: 2019-10-25
QILU ANIMAL HEALTH PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no special canine distemper and parvovirus vaccine for raccoon dogs on the market, and many farmers use alternative vaccines, which cannot achieve better immune protection effect. Therefore, secondary immunization of raccoon dogs is often required, which not only increases the cost, It also increases the workload of the farmers, and at the same time causes a greater stress response in the raccoon dogs
Moreover, most canine distemper vaccines are live vaccines, and there is a risk of strong virulence; the vaccines prepared for parvovirus enteritis are mainly inactivated whole virus vaccines or live vaccines. Inactivated vaccines have poor immunogenicity, and live vaccines have virulence. The risk of returning to strength

Method used

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  • Raccoon dog parvoviral enteritis and canine distemper combined inactivated vaccine and preparation method thereof
  • Raccoon dog parvoviral enteritis and canine distemper combined inactivated vaccine and preparation method thereof
  • Raccoon dog parvoviral enteritis and canine distemper combined inactivated vaccine and preparation method thereof

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Experimental program
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Effect test

preparation example Construction

[0076] 1) Preparation of raccoon parvovirus VP2 protein: select vigorously growing Sf9 cells to inoculate cell culture tanks for full suspension culture, and the inoculation density is 1.5×10 6 ~2.0×10 6 cells / ml, cultured at 27°C, when the density of Sf9 cells in the cell culture tank reaches 1.5×10 6 ~3.0×10 6 When cells / ml, inoculate the recombinant baculovirus RDPV-SN1 strain according to MOI=0.2~2.0, culture at 27°C for 72~96h, harvest the cell culture, add 2% BEI solution to it, make the final concentration of BEI be 0.1%, After mixing, inactivate at 37°C for 48 hours, then add 50% sodium thiosulfate solution to the cell culture to make the final concentration 0.2%, and stop the inactivation after stirring for 1 hour; after the sterility and inactivation test, you can get Raccoon parvovirus VP2 protein, hemagglutination titer ≥ 1:2048 for 1% porcine erythrocytes, stored at 2-8°C for later use;

[0077] 2) Preparation of canine distemper virus CDV-11 strain inactivated...

Embodiment 1

[0088] Embodiment 1——Research on suspension culture technology of recombinant baculovirus RDPV-SN1 strain

[0089] 1. Research on culture technology of recombinant baculovirus RDPV-SN1 strain Erlenmeyer flask

[0090] (1) Effect of cell density on hemagglutination titer of expressed protein

[0091] When the Sf9 cell density is 1.5×10 6 cells / ml, 2.0×10 6 cells / ml, 2.5×10 6 cells / ml, 3.0×10 6 Cells / ml, the recombinant baculovirus RDPV-SN1 strain was inoculated according to MOI=0.5, samples were taken at 60h, 72h, 84h, 96h, 108h, and 120h after inoculation, and the hemagglutination titer of the expressed protein was determined.

[0092] Results The recombinant baculovirus RDPV-SN1 strain was inoculated into Sf9 cells with different cell densities, and the density of Sf9 cells was 1.5×10 6 , 2.0×10 6 , 2.5×10 6 , 3.0×10 6 There was no significant difference in the hemagglutination titer of the expressed protein at the time of cells / ml, and the hemagglutination titer of t...

Embodiment 2

[0113] Embodiment 2——Canine distemper virus CDV-11 strain suspension culture technique research

[0114] 1. Study on the culture technology of canine distemper virus CDV-11 strain 7L cell culture tank

[0115] (1) Microcarrier Preparation and Sterilization 2+ , Mg 2+ Wash with PBS (0.1mol / L, pH value 7.2, the same below) for 2 to 3 times, add PBS and sterilize at 121°C for 30min. Discard PBS after sterilization, add MEM cell culture medium containing 8% newborn bovine serum, and store at 36.5°C, containing 5% CO 2 Place in the incubator for 4h.

[0116] (2) Vero cell microcarrier suspension culture selects Vero spinner bottle cells with good shape and vigorous growth, digests with EDTA-trypsin dispersion to prepare cell suspension, inoculates agitated cell culture flask, the rotating speed is 30r / min, and the cell density is 4.5×10 5 cells / ml, the microcarrier concentration is 4g / L, and the culture temperature is 36.5℃~37.5℃.

[0117] (3) Effect of culture temperature on...

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Abstract

The invention relates to a raccoon dog parvoviral enteritis and canine distemper combined inactivated vaccine and a preparation method thereof. The raccoon dog parvoviral enteritis and canine distemper combined inactivated vaccine comprises the effective ingredients of mink parvoviral VP2 protein, acquired by culturing recombinant rhabdovirus RDPV-SN1 transfected Sf9 cells, and a canine distempervirus inactivated antigen. In order to solve the problem that the racoon dog and canine distemper and parvovirus combined active vaccine has the risk of virulence return, while the combined inactivated vaccines used in the market generally cause low immunity after the raccoon dog parvovirus is inactivated, the invention provides a combined inactivated vaccine prepared with raccoon dog parvoviral VP2 protein and the canine distemper virus inactivated antigen having high viral content. The combined inactivated vaccine herein is applicable to the prevention of raccoon dog parvoviral enteritis andcanine distemper, has the advantages of good immunogenicity, high safety, high controllability, high culture efficiency, high automation level, large-scale production and the like, can provide doublepreventions and has a good application prospect.

Description

technical field [0001] The invention relates to a dual inactivated vaccine of raccoon parvovirus enteritis and canine distemper and a preparation method thereof, belonging to the field of veterinary biological products. Background technique [0002] Raccoon parvovirus enteritis is an acute and highly contagious infectious disease caused by raccoon parvovirus of the family Parvoviridae. At present, raccoon dog parvovirus enteritis widely exists in various countries in the world. The disease mainly causes enteritis of raccoon dogs, and the mortality rate can reach 30% to 60%. [0003] Canine distemper is an acute, febrile, and highly contagious infectious disease caused by canine distemper virus of the genus Morbillivirus in the Paramyxoviridae family. Most of the young animals have an acute and fatal course, and adult animals can become chronic and persistent. Infection, the clinical symptoms are mainly characterized by biphasic body temperature rise, accompanied by catarrhal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/23A61K39/175A61P31/20A61P31/14C07K14/015C12N15/866C12N15/35
CPCA61K39/12A61P31/20A61P31/14C07K14/005C12N15/86A61K2039/552A61K2039/70A61K2039/5252A61K2039/55505C12N2750/14334C12N2760/18434
Inventor 董海曼李雯宋晓飞葛平萍秦旭伟李营贾爱琴王蕾徐龙涛
Owner QILU ANIMAL HEALTH PROD
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