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Canine distemper attenuated strain as well as vaccine composition prepared from same and application

A vaccine composition and canine distemper virus technology, applied in the directions of vaccines, veterinary vaccines, biochemical equipment and methods, etc., can solve the problems such as the canine distemper epidemic cannot be effectively controlled, the antigen targeting is not strong, and achieve good Protective effect, blocking continuous infection, good immunogenic effect

Active Publication Date: 2021-03-19
PU LIKE BIO ENG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These traditional CDV vaccine strains have a large gap in genotype with the CDV wild strains that have been popular in China for the past 10 years, and have the defect of poor antigenicity. Especially in recent years, canine distemper virus has continuously mutated and infected hosts have continued to expand. As a result, the domestic canine distemper epidemic has been unable to be effectively controlled, and it is of great practical significance to develop an attenuated vaccine that is safe and has good immunogenicity

Method used

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  • Canine distemper attenuated strain as well as vaccine composition prepared from same and application
  • Canine distemper attenuated strain as well as vaccine composition prepared from same and application
  • Canine distemper attenuated strain as well as vaccine composition prepared from same and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The acquisition of embodiment 1 canine distemper virus HL001-M3 strain

[0035] 1. get the well-grown Vero cells, digest with trypsin, inoculate in the cell bottle, with the cell growth liquid ( Adjust the pH to 6.8-7.2) and continue culturing at 33°C-37°C to form a good monolayer for virus inoculation.

[0036] 2. Canine distemper virus HL001 strain is inoculated with the above-mentioned passaging cell monolayer that grows well, with the cell growth solution (pH Adjusted to 6.8-7.2) Continue culturing at 33°C-37°C, and after 72h-96h, when more than 80% of the cells have lesions, harvest the cell culture virus liquid.

[0037]3. Repeat the above steps for continuous subculture to obtain the attenuated strain of canine distemper virus, and sequence the obtained attenuated canine distemper virus strain. The results show that the 1430 nucleotide of the H gene undergoes a stable mutation from A→T, resulting in H protein H477L Amino acid variation, 1556 nucleotides have a ...

Embodiment 2

[0038] Example 2 Study on Biological Characteristics of Canine Distemper Virus HL001-M3 Strain

[0039] 1. Pathogenicity test

[0040] Fifteen healthy susceptible antigen-negative dogs aged 2 to 3 months were randomly divided into 3 groups with 5 dogs in each group. See Table 1 for the grouping and challenge.

[0041] Table 1 Grouping of pathogenic test animals

[0042] group Inoculation strain Inoculation dose 1 HL001-M3 strain Nasal drops 2ml, intraperitoneal 4ml (10 5.5 FAID 50 / ml)

2 HL001 strain Nasal drops 2ml, intraperitoneal 4ml (10 5.5 FAID 50 / ml)

3 DMEM medium Nasal drops 2ml, intraperitoneal 4ml

[0043] Observed for 21 days after virus inoculation, and observed and recorded the dog's body temperature, energy, appetite, feces, and eye and nose secretions every morning and evening. The specific results are shown in Table 2.

[0044] Table 2 Pathogenicity of HL001-M3 strain to dogs

[0045]

[0046] The resul...

Embodiment 3

[0072] Preparation of Example 3 Canine Distemper Virus HL001-M3 Strain Attenuated Live Vaccine

[0073] 1. Proliferation of virus

[0074] The canine distemper virus HL001-M3 strain virus seed prepared in Example 1 was inoculated into the Vero cell suspension synchronously, added DMEM culture fluid containing 2% newborn calf serum, and placed at 37° C., 5% CO 2 nourish. After 80% of the cells were damaged, the virus was harvested, the virus titer was measured, and stored at low temperature.

[0075] 2. Preparation of protective agent

[0076] Add 40 g of sucrose and 8 g of gelatin per 100 ml of deionized water, and after fully melting, sterilize with high pressure steam (121° C. for 30 min).

[0077] 3. Vaccine Preparation

[0078] The virus liquid prepared and preserved above is mixed with the protective agent at a ratio of 1:1 (volume ratio), and freeze-dried. The specific ratio of vaccine content is shown in Table 4.

[0079] Table 4 Canine distemper virus HL001-M3 st...

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Abstract

The invention provides a canine distemper virus attenuated strain HL001-M3 which has good immunogenicity, genetic stability and low toxicity, and a prepared live vaccine can prevent and treat canine distemper, protect epidemic strains and protect various animals.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to an attenuated canine distemper strain and the prepared vaccine composition and application thereof. Background technique [0002] Canine distemper virus (CDV) is a single-stranded RNA virus belonging to the Morbillivirus genus of the family Paramyxoviridae, and the virus particles are round or shapeless. In recent years, with the change of the ecological environment and the evolution of animals and viruses, the natural infection hosts of CDV have been expanded from the traditional Canidae (including dogs, foxes, raccoon dogs, etc.), Procyonidae and Weaselidae (mink, etc.) to All 8 families of Carnivora, Artiodactyla Suidae, Primate Macaca and Pinniped Sealidae and other animals, and they are showing a trend of continuous expansion. [0003] Canine distemper (Canine distemper, CD) is an acute, febrile, highly contagious infectious disease caused...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/04A61K39/175A61P31/14C12R1/93
CPCC12N7/00A61K39/12A61P31/14C12N2760/18421C12N2760/18434A61K2039/5254A61K2039/552
Inventor 田克恭刘玉秀习向锋张许科
Owner PU LIKE BIO ENG
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