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Canine parvovirus attenuated strain, vaccine composition prepared from canine parvovirus attenuated strain and application

A technology of canine parvovirus and vaccine composition, which is applied in the direction of viruses, vaccines, antiviral agents, etc., and can solve problems such as harm, loss, and inability to protect dogs in the breeding industry

Active Publication Date: 2021-07-06
PU LIKE BIO ENG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Among the currently commercialized canine distemper and parvovirus dual-combination live vaccines, the canine distemper virus vaccine strains (Onderstepoort strain, Snyder Hill strain) are American-1 type, while the domestically prevalent Asian-1 type is the same. The parvovirus vaccine strains (NL-35-D strain, 154 strain) are all early CPV-2 types. With the continuous evolution of canine parvovirus, CPV-2 strains have been replaced by mutant strains new CPV-2a, New CPV-2b and CPV-2c are replaced, but the new CPV-2a is mainly popular in China at present, and the new CPV-2a strain that is popular in China is different from the CPV-2 strain at the nucleotide and amino acid levels There are differences and cannot effectively protect dogs
[0006] 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 parvovirus attenuated strain, vaccine composition prepared from canine parvovirus attenuated strain and application
  • Canine parvovirus attenuated strain, vaccine composition prepared from canine parvovirus attenuated strain and application
  • Canine parvovirus attenuated strain, vaccine composition prepared from canine parvovirus attenuated strain and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 The acquisition of canine parvovirus JM35 strain

[0043] 1. Take well-grown cat kidney cells (F81 cells), digest them with trypsin, and inoculate them in cell flasks with 90% to 98% volume ratio of RPMI 1640 culture medium and 2% to 10% volume ratio of newborn bovine Serum cell growth solution (pH adjusted to 6.8-7.2) was cultured at 33°C-37°C for virus inoculation.

[0044] 2. The canine parvovirus JM strain was inoculated synchronously with the above passaged cells, and the cell growth liquid (pH adjusted to 6.8-7.2 ) at 33° C. to 37° C. to continue culturing, and after 72 h to 120 h, when more than 80% of the cells have pathological changes, the cell culture virus liquid is harvested.

[0045] 3. Repeat the above steps for continuous subculture to obtain the attenuated canine parvovirus strain, and sequence the obtained attenuated canine parvovirus strain. The results show that the 63rd nucleotide of the VP2 gene undergoes a stable synonymous mutation fro...

Embodiment 2

[0046] Example 2 Study on Biological Characteristics of Canine Parvovirus JM35 Strain

[0047] 1. Pathogenicity test

[0048] 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.

[0049] Table 1 Grouping of test animals for pathogenicity of canine parvovirus JM35 strain

[0050] group Inoculation strain Inoculation dose 1 JM35 strain Subcutaneous injection of 4ml (10 6.5 FAID 50 / ml)

2 JM strain Subcutaneous injection of 4ml (10 6.5 FAID 50 / ml)

3 RPMI 1640 Medium Subcutaneous injection 4ml

[0051] Observed for 10 days after virus inoculation, and observed and recorded the clinical manifestations of dogs such as spirit, appetite, and feces every day. The specific results are shown in Table 2.

[0052] Table 2 Pathogenicity of canine parvovirus JM35 strain to dogs

[0053]

[0054] The results sho...

Embodiment 3

[0080] Embodiment 3 Preparation of Canine Parvovirus JM35 Strain Attenuated Live Vaccine

[0081] 1. Proliferation of virus

[0082]The canine parvovirus JM35 strain virus seed prepared in Example 1 was synchronously inserted into the F81 cell suspension, and the RPMI 1640 culture solution containing 90% to 98% by volume and 2% to 10% by volume of newborn bovine serum were added for cell growth solution (pH adjusted to 6.8-7.2) was cultivated at 33°C-37°C. When the cytopathic rate reaches about 80%, the virus is collected by freezing and thawing, and the virus content is measured, and stored at low temperature.

[0083] 2. Preparation of protective agent

[0084] 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).

[0085] 3. Vaccine Preparation

[0086] The virus liquid prepared and preserved above is mixed with the protective agent at a ratio of 1:1 (volume ratio), and fre...

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Abstract

The invention provides a canine parvovirus attenuated strain JM35 with a deposit number of CCTCC NO.V201965. The invention further provides a vaccine composition containing an immune dose of the canine parvovirus attenuated strain JM35 or a culture thereof. The vaccine composition not only can completely prevent and protect canine parvoviruses of different serotypes and canine parvoviruses of different regional sources, but also can block continuous infection of the canine parvoviruses.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, and in particular relates to attenuated canine parvovirus strains, vaccine compositions prepared therefrom and applications thereof. Background technique [0002] Canine distemper virus (CDV) is a viral disease with high morbidity and mortality worldwide. With the change of ecological environment and the evolution of animals and viruses, its host has been changed from traditional Canidae (including dogs, foxes, raccoons, etc.), Procyonidae and Mustelidae (mink, etc.) animals have expanded to all 8 families of Carnivora and many animals such as Artiodactyla Suidae, Primates Rhesus and Pinnipeds Sealidae , and the host range of CDV presents a trend of continuous expansion. [0003] Canine parvovirus (Canine Parvovirus, CPV) was first discovered in 1978 and is currently prevalent all over the world. Canine parvovirus is the pathogen that causes acute hemorrhagic enteritis in dogs and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/23A61P31/20A61K39/175A61P31/14C12R1/93
CPCC12N7/00A61K39/12A61P31/20A61P31/14C12N2750/14321C12N2750/14351C12N2750/14334A61K2039/5254A61K2039/552A61K2039/70C12N2760/18434
Inventor 田克恭吴洪超张许科
Owner PU LIKE BIO ENG
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