Bivalent live vaccine against canine distemper and parvovirus diseases, and preparation method thereof

A technology of canine distemper virus and canine parvovirus, applied in biochemical equipment and methods, antiviral agents, virus/bacteriophage, etc., can solve the problems of low protection rate and weak immunogenicity of virus strains, and achieve immunity Strong originality, suitable for industrial production, and the effect of controllable quality

Active Publication Date: 2017-02-15
兆丰华生物科技(南京)有限公司 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] According to incomplete statistics, the combined infection rate of canine distemper and canine parvovirus is about 10%, while

Method used

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  • Bivalent live vaccine against canine distemper and parvovirus diseases, and preparation method thereof
  • Bivalent live vaccine against canine distemper and parvovirus diseases, and preparation method thereof
  • Bivalent live vaccine against canine distemper and parvovirus diseases, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of poisonous seeds for production

[0029] Preparation of Canine Distemper Virus BJ / 120 Strain Production Virus Seed

[0030] Canine distemper virus CDV / BJ / 120 strain was inoculated on Vero cell monolayer at 2%, cultured at 33°C for 96-120 hours, and harvested when the cytopathy reached 80%, and after freezing and thawing at -20°C for 3 times, the virus was sampled Content determination, sterility test, mycoplasma test and exogenous virus test, quantitative subpackaging after passing the identification, freeze-drying or storage below -40°C.

[0031] Preparation of Canine Parvovirus CPV / FJ / 58 Strain Production Virus Seeds

[0032]Canine parvovirus CPV / FJ / 58 strain was inoculated in DK cells with 2%, and the cells grew into a single layer after 24-48 hours, then replaced with MEM nutrient solution containing 2% bovine serum, cultured at 37°C for 96-120 hours, harvested, and transferred After freezing and thawing at -20°C for 3 times, samples were t...

Embodiment 2

[0033] Example 2 Preparation of virus liquid for preparation

[0034] Preparation of Canine Distemper Virus Liquid

[0035] Select a well-grown Vero cell monolayer, discard the nutrient solution, add 2% canine distemper virus to the MEM nutrient solution containing 2% bovine serum, culture at 33°C, and observe the cell lesions every day, when the cell lesions reach 80% Harvest at around time, freeze and thaw three times at -20°C, and store the virus liquid below -40°C.

[0036] Preparation of Canine Parvovirus Liquid

[0037] Select well-growing DK cells, disperse them with trypsin, add canine parvovirus seeds to the cell solution at a ratio of 2%, and wait for 24 to 48 hours. After the cells form a single layer, replace them with MEM containing 2% bovine serum Nutrient solution, continue to cultivate at 37°C for 96 to 120 hours, transfer the culture bottle to -20°C to freeze and thaw three times, and then freeze the virus solution below -40°C.

Embodiment 3

[0038] Example 3 Preparation of Canine Distemper and Parvovirus Dual Live Vaccine

[0039] Mix the canine distemper virus liquid prepared in Example 2 and the canine parvovirus liquid in a volume ratio of 1:1, and then mix it with the freeze-drying protective agent in a volume ratio of 1:1, shake well, and quantitatively dispense in In the vaccine bottle, the freeze-drying procedure is -40°C for 6 to 7 hours, -10°C for 16 hours, 30°C for 6 hours, capping and labeling.

[0040] The freeze-drying protective agent is: 8g sucrose, 5g gelatin, 2.5g mannitol, 2g trehalose, 0.3 mannitol, 0.5g arginine, 100ml deionized water, adjust the pH value to 7.2-7.6; Bacteria for 30 minutes.

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Abstract

The invention discloses a bivalent live vaccine against canine distemper and parvovirus diseases, and a preparation method thereof. The bivalent live vaccine contains an attenuated anti-canine distemper vaccine strain with a microbial accession number of CGMCC No. 10980 and an attenuated anti-parvovirus distemper vaccine strain with a microbial accession number of CGMCC No. 7408. According to the invention, a canine distemper virus and a canine parvovirus separated from wild viruses of diseased Chinese dogs and having undergone attenuation via domestication are used for preparation of a bivalent vaccine together, and a freeze-drying protective agent is added so as to prepare the bivalent live vaccine against canine distemper and parvovirus diseases. The bivalent live vaccine is reliable in security, good in immune effect, strong in targeting performance, suitable for prevention of diseased caused by diseased Chinese dogs and applicable to prevention of canine distemper and parvovirus diseases.

Description

technical field [0001] The invention relates to a veterinary dual live vaccine, in particular to a canine distemper and canine parvovirus dual live vaccine and a preparation method thereof, belonging to the field of veterinary dual vaccines. Background technique [0002] Canine distemper and canine parvovirus are two severe infectious diseases that seriously harm the dog industry in my country. [0003] Canine distemper (CD) is a highly contagious infectious disease caused by canine distemper virus (CDV), with various clinical symptoms, and is prone to secondary mixed infection and secondary infection of other bacteria and viruses , the mortality rate can be as high as 80%, and the sick animals are prone to sequelae such as paralysis, convulsions, and epileptic seizures after recovery. The disease was discovered worldwide in 1926, endangering my country's dog industry, fur animal breeding industry and wild animal protection industry, causing a large number of dogs, mink, fo...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/23A61K39/175A61P31/14A61P31/20C12R1/93
Inventor 王海燕廖丽晶赵锐刘飞马金枝王贵华赵亚荣
Owner 兆丰华生物科技(南京)有限公司
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