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Canine parainfluenza virus strain and application thereof

A technology of canine parainfluenza virus and canine parvovirus, which is applied in the direction of viruses, antiviral agents, and viral antigen components, can solve the problems of strong virulence, low immune efficacy of inactivated vaccines, and inability to completely resist infection. The effect of good biological security, good prevention and control effect

Active Publication Date: 2018-05-11
PU LIKE BIO ENG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Vaccine is the main measure to prevent and control the disease. As early as 1970, Emay et al. developed the CPIV D-008 attenuated vaccine, which can produce neutralizing antibodies by intramuscular or subcutaneous inoculation. The use of this vaccine can reduce the incidence of the disease and relieve clinical symptoms. Symptoms, but cannot fully fight off infection
At present, many companies at home and abroad are providing vaccines containing CPIV, but in clinical practice, it is found that some CPIV-positive dogs have been strictly vaccinated with CPIV vaccine history
On the other hand, the attenuated vaccine has the risk of virulence returning to strong and loose virus during use, while the virulence of the existing isolated virulent strains is significantly attenuated during the cultivation process, and the immune efficacy of the prepared inactivated vaccine is not high. There are related inactivated vaccine products appearing

Method used

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  • Canine parainfluenza virus strain and application thereof
  • Canine parainfluenza virus strain and application thereof
  • Canine parainfluenza virus strain and application thereof

Examples

Experimental program
Comparison scheme
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Embodiment approach

[0028] As an embodiment of the present invention, the inactivating agent includes but not limited to β-propiolactone BPL, binary ethyleneimine BEI, formalin, formaldehyde, N-acetylethyleneimine AEI, polyhexamethylene hydrochloride Methylguanidine PHMG.

[0029] As an embodiment of the present invention, in the vaccine composition of the present invention, the culture of the canine parainfluenza virus strain is a culture cultured for 1-18 generations.

[0030] As an embodiment of the present invention, in the vaccine composition of the present invention, the culture of the canine parainfluenza virus strain is a culture cultured for 6-18 generations.

[0031] As an embodiment of the present invention, in the vaccine composition of the present invention, the adjuvant includes: (1) aluminum gel adjuvant, saponin, avridine, DDA; (2) water-in-oil emulsion, water Oil-in-emulsions, water-in-oil-in-water emulsions; or (3) polymers of acrylic or methacrylic acid, copolymers of maleic a...

Embodiment 1

[0049] The separation and identification of embodiment 1 canine parainfluenza virus

[0050] The wild strain of CPIV was isolated from clinically infected animals with canine parainfluenza virus. The clinical symptoms of the infected animals were: mental depression, fever, a large amount of mucus from the nasal cavity, opaque secretions, dry cough, etc., which were used as the criteria for the selected animals. The lung tissue of the infected animal was taken and ground with a tissue homogenizer to obtain a tissue homogenate.

[0051] Dilute the lung tissue homogenate of sick dogs to 10% W / V suspension with serum-free DMEM culture fluid, filter and sterilize with a 0.22 μm filter membrane, and inoculate the filtrate into Vero monolayer cells (purchased from Shanghai Enzyme Detection Technology Co., Ltd.) for virus isolation, cell fusion lesion appeared in 24 to 48 hours, and the cell lesion reached more than 80% in about 96 hours, and the freeze-thawed virus was collected as t...

Embodiment 2

[0052] Embodiment 2 canine parainfluenza virus clone

[0053] 1. Sequencing of the whole genome of canine parainfluenza virus

[0054] Referring to the CPIV5 sequence (JQ743318.1) submitted in GenBank, 13 pairs of primers were designed. The amplified fragments partially overlap with each other and can be connected to cover the whole genome. The primer sequences are shown in Table 1.

[0055] Table 1 Sequence list of canine parainfluenza virus whole genome sequencing primers

[0056]

[0057]

[0058] Referring to the instructions of the RNA extraction kit, extract the total RNA of the CPIV virus liquid isolated in Example 1, and use the downstream primers of P1 to P13 to perform reverse transcription reactions respectively to prepare the cDNA of the virus. Using the obtained cDNA as a template, use the designed 13 pairs of primers amplified the whole genome of the virus. After the PCR product was identified by 1% agarose gel electrophoresis, the size of the band was in...

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Abstract

The invention relates to a canine parainfluenza virus strain and application thereof. The canine parainfluenza virus HeN0718 strain has strong toxicity and good immunogenicity. A vaccine composition prepared from an inactive antigen of the canine parainfluenza virus HeN0718 strain can immunize an animal to allow the animal to rapidly produce an antibody. The vaccine composition can be effective in prevention and / or treatment of current canine parainfluenza related diseases. The vaccine composition is safe and can be combined with other antigens.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a canine parainfluenza virus strain and a vaccine composition, a preparation method and an application thereof. Background technique [0002] Canine Parainfluenza virus (CPIV) is a member of Paramyxoviridae, Paramyxoviridae, and Mumps virus. It is a single-stranded negative-sense RNA virus that is not segmented. The shape of CPIV particles is variable under the electron microscope. Generally round, with a diameter of 80nm to 200nm, but often irregular in shape due to capsule damage, showing pleomorphic virus particles. [0003] CPIV has a wide host spectrum, including dogs, guinea pigs, hamsters, mice, tigers, raccoon dogs, minks, etc. It often causes bronchitis and bronchial pneumonia in dogs and damages respiratory epithelial cells. After dogs are infected with CPIV, the virus can be detected from respiratory secretions. The virus particles c...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/155A61K39/295A61K39/175A61K39/23A61P31/14A61P31/20C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/70C12N7/00C12N2750/14034C12N2760/18434C12N2760/18621C12N2760/18634
Inventor 田克恭刘玉秀刘彩红孙进忠张许科
Owner PU LIKE BIO ENG
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