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Canine parainfluenza virus attenuated strain, application thereof and vaccine

A technology of canine parainfluenza virus and attenuated strains, which is applied to veterinary vaccines, viruses, vaccines, etc., can solve the problems of high cost and weak pertinence, and achieve the effect of strong pertinence, low production cost and high virus price

Active Publication Date: 2021-06-04
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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  • Canine parainfluenza virus attenuated strain, application thereof and vaccine
  • Canine parainfluenza virus attenuated strain, application thereof and vaccine
  • Canine parainfluenza virus attenuated strain, application thereof 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 material: A puppy from Changchun Pet Hospital in Jilin Province had 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. Collect five parts of nasal secretion disease materials (No. 1, No. 2, No. 3, No. 4, No. 5) in total, take their nasal secretions and add them into 3 times PBS buffer solution containing penicillin and streptomycin (pH 7.2) After repeated freezing and thawing for 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 the bacterial test is negative.

[0047] Cells: Vero (F87) cell line was purchased from China Veterinary and Veterinary Drug Control Institute, and was preserved and suppli...

Embodiment 2

[0058] Example 2 Comprehensive Identification of Canine Parainfluenza Attenuated Strain CPIV / 15

[0059] Viral nucleic acid type identification: 5-iododeoxyuridine (5-IUDR) method: culture Vero cells in a 96-well plate, and after the cells grow into a monolayer, pour out the growth medium and replace it 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, and then inoculate different dilutions of the virus solution in the cell culture wells, each dilution was repeated 4 times, and observed daily after inoculation Cytopathy. Take the negative logarithm of the highest dilution factor of the CPE of the virus as the titer of the virus, and determine the TCID of the virus growing on Vero cells 50 At the same time, the culture solution 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, and th...

Embodiment 3

[0092] Example 3 Dog Body Passage Reversion Test

[0093] Test materials and methods:

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

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

[0096] Experimental animals: The dogs used in the experiment are 42-49 days old healthy beagle dogs with CPIV antibody SN negative or ≤2 that have not been immunized with canine parainfluenza virus vaccine.

[0097] Cell subculture stability test: use Vero subculture cells, adopt the method of synchronous poisoning, 37°C 5% CO 2 Cultivate under conditions, cultivate for 5 to 7 days and more than 80% of the cytopathic changes (CPE) will be harvested, and then continue to use Vero to culture and pass down to 60 generations (CPIV / 15F60) to observe the changes of each g...

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Abstract

The invention relates to the field of veterinary drugs, in particular to a canine parainfluenza virus attenuated strain, application thereof and a vaccine. The invention provides the canine parainfluenza virus attenuated strain with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.19993. The canine parainfluenza virus attenuated strain disclosed by the invention is strong in pertinence, can be used for dogs of various varieties and various ages and also can be used for pregnant dogs, and the generated antibody is high and the immune period is long.

Description

technical field [0001] The invention relates to the field of veterinary medicine, in particular to an attenuated strain of canine parainfluenza virus, its application and a vaccine. Background technique [0002] Canine parainfluenza virus (CPIV) is the main pathogen of infectious diseases that cause canine upper respiratory tract infection and encephalomyelitis. Filaments up to several microns, with a diameter of about 80nm to 200nm. Canine parainfluenza virus has a strong infectivity to the nervous system of dogs, foxes, and raccoon dogs. Severe illness can lead to death, which seriously affects the development of dog and fox breeding industries. Canine parainfluenza virus can infect, especially in puppies. The main way of infection is the respiratory tract, and the respiratory secretions can infect other dogs through air and dust, and can also be transmitted through direct contact. During the infection period, the dog developed other bacterial infections due to decrease...

Claims

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Application Information

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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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