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Attenuated strain of canine parainfluenza virus, its application and vaccine

A technology for canine parainfluenza virus and canine parainfluenza, applied in veterinary vaccines, viruses, vaccines, etc., can solve the problems of high cost and weak pertinence, and achieve the effects of strong pertinence, low production cost and simple production process

Active Publication Date: 2022-07-29
BEIJING HUAXIA XINGYANG BIOLOGICAL SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to prevent such infectious diseases, the current vaccines at home and abroad have shortcomings such as weak pertinence and high cost

Method used

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  • Attenuated strain of canine parainfluenza virus, its application and vaccine
  • Attenuated strain of canine parainfluenza virus, its application and vaccine
  • Attenuated strain of canine parainfluenza virus, its application and vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Isolation and identification of natural attenuated CPIV / 15 strain of canine parainfluenza virus.

[0045] Materials and methods

[0046] Source of disease materials: From the pet hospital in Changchun City, Jilin Province, the puppy developed serous or mucopurulent rhinorrhea around the nostrils, accompanied by clinical symptoms such as mild cough, and was initially diagnosed as suspected canine parainfluenza virus disease. A total of five nasal secretion disease materials (numbered as No. 1, No. 2, No. 3, No. 4, No. 5) were collected, and their nasal secretions were added to 3 times PBS buffer (pH) containing penicillin and streptomycin. 7.2) After repeated freezing and thawing 3 times, centrifuge at 8000r / min at 4°C for 15min, filter and sterilize with a sterile filter (0.22μm), and store at -30°C after negative bacterial test.

[0047] Cells: Vero (F87) cell line was purchased from China Veterinary and Veterinary Drugs Supervision Institute, and kept and s...

Embodiment 2

[0058] Example 2 Comprehensive identification of canine parainfluenza attenuated strain CPIV / 15

[0059] Identification of viral nucleic acid types: 5-Iododeoxyuridine (5-IUDR) method: Vero cells were cultured in a 96-well plate, and after the cells grew into a monolayer, the growth medium was poured out and replaced with 2% fetal bovine serum DMEM maintenance solution, and 5-IUDR was added to the maintenance solution to make the final concentration 50 μg / mL, then inoculated with different dilutions of virus solution in the cell culture wells, each dilution was repeated 4 times, and observed daily after inoculation cytopathic. The TCID of the virus grown on Vero cells was determined by taking the negative logarithm of the highest dilution at which the virus appeared CPE as the virus titer 50 Changes, while the culture medium without 5-IUDR was used as a control. Because 5-IUDR has obvious inhibitory effect on DNA virus, it has no or only low inhibitory effect on RNA virus, a...

Embodiment 3

[0092] Example 3 Canine Passaging Reversion Test

[0093] Test materials and methods:

[0094] Cells: African green monkey kidney passage cells (Vero), purchased from China Veterinary Drug Administration, and preserved by Beijing Shengtaier Technology Co., Ltd.

[0095] Virus species: Canine parainfluenza virus attenuated CPIV / 15 strains, F50 generation; preserved and provided by Beijing Shengtaier Technology Co., Ltd.

[0096] Experimental animals: The experimental dogs were 42- to 49-day-old healthy beagle dogs with CPIV antibody SN negative or ≤ 2 who had not been immunized with canine parainfluenza virus vaccine.

[0097] Cell passage stability test: use Vero passage cells, adopt the method of simultaneous inoculation, 37 ℃ 5% CO 2 Cultivated under the condition of culture, more than 80% cytopathic effect (CPE) occurred in 5 to 7 days, harvested, and then continued to use Vero culture to subculture to 60 generations (CPIV / 15F60) to observe the changes of cells in each ge...

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Abstract

The invention relates to the field of veterinary medicine, in particular to an attenuated strain of canine parainfluenza virus, an application thereof and a vaccine. The present invention proposes an attenuated strain of canine parainfluenza virus, and the deposit number is CGMCC No.19993. The canine parainfluenza virus attenuated strain of the invention has strong pertinence, can be used for dogs of various breeds and ages, and can also be used for pregnant dogs, with high antibody production and long immune period.

Description

technical field [0001] The invention relates to the field of veterinary medicine, in particular to an attenuated strain of canine parainfluenza virus, an application thereof and a vaccine. Background technique [0002] Canine parainfluenza virus (CPIV) is the main pathogen of infectious diseases that cause upper respiratory tract infection and encephalomyelitis in dogs. The nucleic acid type of CPIV is single-stranded negative-stranded RNA, polymorphic, generally spherical, and long Filaments up to several micrometers, about 80nm to 200nm in diameter. Canine parainfluenza virus has strong infectivity to the nervous systems of dogs, foxes and raccoon dogs. Severe illness can lead to death, thus seriously affecting the development of breeding industries such as dogs and foxes. Canine parainfluenza virus can be infected, especially in puppies. The main route of infection is the respiratory tract, and the respiratory secretions infect other dogs through the air and dust, and c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N7/08A61K39/295A61K39/155A61K39/235A61K39/23A61K39/175A61K39/215A61P31/16A61P31/20A61P31/14C12R1/93
CPCC12N7/00A61K39/12A61P31/16A61P31/20A61P31/14C12N2760/18621C12N2760/18634C12N2760/18664C12N2710/10034C12N2750/14034C12N2760/18434C12N2770/20034A61K2039/552A61K2039/5254A61K2039/70
Inventor 金扩世郭智伟贺亚奇胡义彬李晓亮李三鹏尚川川商云鹏赵卓王力江厚生
Owner BEIJING HUAXIA XINGYANG BIOLOGICAL SCI & TECH
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