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A canine distemper virus canine parvovirus canine infectious hepatitis virus triple live vaccine

A technology for canine parvovirus and canine distemper virus, which is applied in the directions of viruses, vaccines, antiviral agents, etc., can solve the problems of many passages, difficult to guarantee immunogenicity and safety, and unclear background of virus species.

Active Publication Date: 2021-08-24
LIAONING YIKANG BIOLOGICAL CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, most of these vaccines currently in use are replicas of foreign vaccines. The background of the virus species is unclear and the number of passages on cells is high, so it is difficult to guarantee immunogenicity and safety.

Method used

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  • A canine distemper virus canine parvovirus canine infectious hepatitis virus triple live vaccine
  • A canine distemper virus canine parvovirus canine infectious hepatitis virus triple live vaccine
  • A canine distemper virus canine parvovirus canine infectious hepatitis virus triple live vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Embodiment 1. Acquisition of the original strain of canine distemper virus

[0109] (1) Case: A puppy from a dog farm of a dog professional cooperative in Liaoyang City, Liaoning Province developed the disease. The symptoms were: high fever, depression, nasal discharge and eye mucus, and neurological symptoms in the later stage. Judging by the symptoms, it is suspected to have canine distemper.

[0110] (2) Virus isolation: get the brain tissue of the sick dog, wash the brain tissue with sterilized normal saline for 3 times, add the brain tissue into sterilized cold normal saline at a ratio of 1g tissue: 10ml, put it into a tissue mixer, and wash it with 10000 ~12000r / min fully smashed the brain tissue, repeated freezing and thawing below -15℃ for 3 times, filtered twice with 3 layers of sterilized gauze, collected the filtrate, and centrifuged at 12000r / min at 4℃ for 30 minutes, and collected the supernatant , adding filter-sterilized final concentrations of 1000 IU / m...

Embodiment 2

[0120] Embodiment 2. The domestication of canine distemper virus strain and the test of strain biochemistry, genetic change

[0121] (1) Virus subculture: the cell culture solution a (containing virus strain a) of embodiment 1 is continuously subcultured to 150 generations on Vero cells, and the specific steps are as follows: the Vero cells that have grown into a single layer are discarded and grown solution (composition: DMEM containing 2v / v% newborn bovine serum), inoculate the cell culture solution a of embodiment 1 by 5v / v% inoculation amount, put 33 ℃ containing 5% CO 2 Adsorb in the incubator for 1 hour, add DMEM cell maintenance solution containing 2v / v% newborn calf serum, and place at 33°C with 5% CO 2 Cultivate in an incubator for 5-6 days. When more than 90% of the cells have CPE, the progeny virus is harvested, passed for 150 generations, and the 150th generation is named D2 strain.

[0122] (2) Virus viability test: 120, 135, and 150 generations of viruses were d...

Embodiment 3

[0137] Embodiment 3. The spin bottle culture of canine distemper virus strain (D2 strain)

[0138] (a) Cell expansion culture: expand the cultivated Vero seed cells into a spinner bottle for cultivation, the culture medium is DMEM containing 8v / v% newborn bovine serum, at a temperature of 37°C, the control speed is 10-11 rpm, and the culture is carried out. 24 hours. (b) inoculation: get the well-grown Vero cell monolayer spinner bottle, discard the nutrient solution, and inoculate the canine distemper virus D2 strain (content 10 4.0 TCID 50 / ml), placed at 33°C for adsorption for 1 hour, added DMEM cell maintenance solution containing 2v / v% newborn bovine serum, and placed at 33°C for rotary culture at a speed of 10-11 rpm. (c) Harvesting: observe the cytopathic changes every day, and when the CPE reaches more than 90%, harvest the virus culture medium and freeze and thaw twice at -20°C. At the same time, samples were taken for sterility testing and virus content determina...

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Abstract

The invention discloses a vaccine strain combination for treating, preventing, slowing down or controlling canine distemper, canine parvovirus enteritis and canine infectious hepatitis, comprising: canine distemper virus vaccine with microorganism preservation number CGMCC No.19397 The strain, the canine parvovirus vaccine strain whose microorganism preservation number is CGMCC No.19398 and the canine infectious hepatitis virus vaccine strain whose microorganism preservation number is CGMCC No.19396. The three vaccine strains in the vaccine strain combination have low virulence and good immunogenicity. The invention also discloses a live vaccine composition using the aforementioned vaccine strain combination as an immunogen. The vaccine composition is safe and effective.

Description

technical field [0001] The invention belongs to the field of preventive veterinary medicine, and relates to a method for preparing a combined vaccine of canine distemper virus, canine parvovirus and canine infectious hepatitis virus and its application, in particular to the preparation of a triple live vaccine of canine distemper virus, canine parvovirus and canine infectious hepatitis virus method and its application. Background technique [0002] With changes in the environment and the way humans live and eat, wild animals and companion animals are in closer contact with humans, thus increasing the potential risk of human infection with zoonotic diseases. Dogs are one of the important companion animals for human beings, and they accompany human beings day and night. Therefore, it is imperative to strengthen the prevention and control of major diseases in dogs. Canine distemper, canine parvovirus and canine infectious hepatitis are currently three highly contagious, acute ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/175A61K39/23A61K39/235A61P31/14A61P31/20A61P1/16A61P1/00C12N7/00C12R1/93
CPCA61K39/12A61P31/14A61P31/20A61P1/16A61P1/00C12N7/00A61K2039/70A61K2039/552C12N2760/18451C12N2750/14351C12N2710/10351C12N2760/18434C12N2750/14334C12N2710/10334A61K2039/5254C12N2760/18421C12N2760/18464C12N2750/14321C12N2750/14364C12N2710/10321C12N2710/10364A61K39/23A61K39/29A61K39/175C12N2750/14034
Inventor 李凤艳舒秀伟王博王一平罗文有陈生雷刘艳霞
Owner LIAONING YIKANG BIOLOGICAL CORP LTD
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