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Canine distemper virus attenuated strain, vaccine composition prepared from canine distemper virus attenuated strain and application of vaccine composition

A technology of canine distemper virus and vaccine composition, applied in the field of veterinary biological products, can solve problems such as loss, protection of animals that cannot host, and harm to the breeding industry

Pending Publication Date: 2021-12-24
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are differences in nucleotide and amino acid levels between domestic epidemic strains and commercial vaccine strains, which cannot effectively protect host animals
[0005] In particular, since the introduction of these commercially imported vaccines into China, the clinical cases of canine distemper virus and canine parvovirus infection alone or in combination have not been significantly suppressed or reduced. cause great harm and serious loss

Method used

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  • Canine distemper virus attenuated strain, vaccine composition prepared from canine distemper virus attenuated strain and application of vaccine composition
  • Canine distemper virus attenuated strain, vaccine composition prepared from canine distemper virus attenuated strain and application of vaccine composition
  • Canine distemper virus attenuated strain, vaccine composition prepared from canine distemper virus attenuated strain and application of vaccine composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] The acquisition of embodiment 1 canine distemper virus LT90 strain

[0051] 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.

[0052] 2. the above-mentioned passaged cell monolayer that canine distemper virus LT strain inoculates growth is good, with the cell growth liquid (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.

[0053] 3. Repeat the above steps for continuous subculture to obtain attenuated canine distemper virus strains, and sequence the obtained attenuated canine distemper virus strains. The results show that the amino acids of M protein V140F, A231V and D290E are stably mutated, and the amino acids of H protein H477L and R519I are stably mutated. F protein N209S and L390...

Embodiment 2

[0054] Example 2 Study on Biological Characteristics of Canine Distemper Virus LT90 Strain

[0055] 1. Pathogenicity test

[0056] 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.

[0057] Table 1 Grouping of pathogenic test animals

[0058] group Inoculation strain Inoculation dose 1 LT90 strain Nasal drops 2ml, intraperitoneal 4ml (10 5.5 FAID 50 / ml)

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

3 DMEM medium Nasal drops 2ml, intraperitoneal 4ml

[0059] 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.

[0060] Table 2 Pathogenicity of LT90 strain to dogs

[0061]

[0062]

[0063] The results sh...

Embodiment 3

[0088] The preparation of embodiment 3 canine distemper virus LT90 strain attenuated live vaccine

[0089] 1. Proliferation of virus

[0090] The canine distemper virus LT90 strain virus seed prepared in Example 1 was inoculated into the Vero cell suspension synchronously, added DMEM culture fluid containing 2% neonatal calf serum, and placed at 37° C., 5% CO 2 to cultivate. After 80% of the cells were damaged, the virus was harvested, the virus titer was measured, and stored at low temperature.

[0091] 2. Preparation of protective agent

[0092] 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).

[0093] 3. Vaccine Preparation

[0094] 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.

[0095] Table 4 Content ratio of canine diste...

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Abstract

The invention relates to a vaccine composition. The vaccine composition comprises an immunizing dose of canine distemper virus LT90 strain antigen and a freeze-drying protective agent, the antigen of the canine distemper virus LT90 strain is a live whole virus antigen of the canine distemper virus LT90 strain or a culture of the canine distemper virus LT90 strain, and the preservation number of the canine distemper virus LT90 strain is CCTCC (China Center For Type Culture Collection) NO.V202030. The vaccine composition disclosed by the invention can be used for preventing and treating existing epidemic canine distemper, can be used for treating dogs infected or repeatedly infected with canine distemper, and can be used for preventing and controlling canine distemper in different regions. The invention also provides the canine distemper virus LT90 strain, and a preparation method and application of the vaccine composition.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, and in particular relates to an attenuated strain of canine distemper virus, a prepared vaccine composition and application thereof. Background technique [0002] Canine distemper (Canine distemper, CD) caused by canine distemper virus (CDV) infection is a kind of high morbidity, high mortality and viral disease that exists worldwide. It is characterized by conjunctivitis, catarrhal inflammation of the upper respiratory tract, lungs and gastrointestinal tract, dermatitis, neurological symptoms, and footpad sclerosis, and is easily secondary to mixed infections and secondary infections such as other bacteria and viruses. Seriously affect the health of dogs and other susceptible animals, cause a large number of dogs to die, and can cause serious sequelae of sick animals, causing serious economic losses to pet dogs and the dog industry. [0003] Canine parvovirus (Canine Parvovirus, CP...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K39/23A61K39/165A61K47/26A61K47/42A61P31/14A61P31/20C12N7/00
CPCA61K39/12A61K47/26A61K47/42A61P31/14A61P31/20C12N7/00C12N2760/18451C12N2750/14051C12N2760/18434C12N2750/14034A61K2039/552A61K2039/70A61K2039/5254
Inventor 田克恭刘玉秀张许科
Owner PU LIKE BIO ENG
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