Attenuated vaccine strain of canine adenovirus type 2 and its application

A technology of canine adenovirus and attenuated vaccines, which is applied in the direction of viruses, vaccines, antiviral agents, etc., can solve the problems of poor pertinence and high cost of vaccines, and achieve the effects of long duration of immunity, strong acid resistance, and simple production process

Active Publication Date: 2022-07-26
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 this type of infectious disease, a lot of research is being carried out at home and abroad, but the current vaccines have shortcomings such as poor pertinence and high cost. It is particularly important to develop a new and efficient canine adenovirus type 2 vaccine

Method used

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  • Attenuated vaccine strain of canine adenovirus type 2 and its application
  • Attenuated vaccine strain of canine adenovirus type 2 and its application
  • Attenuated vaccine strain of canine adenovirus type 2 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Isolation and identification of canine adenovirus natural attenuated CAV2 / 15 strain.

[0045] Virus isolation: Most of the weaned puppies from a dog farm in Shenyang City, Liaoning Province had clinical symptoms such as mild cough and serous nasal discharge, and were diagnosed as suspected canine adenovirus disease. There were no obvious pathological changes in anatomy. Tonsils, lungs and hilar lymph nodes were collected from dogs to isolate the virus.

[0046] Cells: Canine kidney passaged cells MDCK (passage 87) cell line was purchased from China Veterinary Drug Administration, and kept and supplied by Beijing Shengtaier Technology Co., Ltd.

[0047]Experimental animals: 42 to 49-day-old healthy beagle dogs that were not immunized with canine adenovirus vaccine and were negative for antigen and antibody were purchased from Shenyang Kangping Laboratory Animal Research Institute.

[0048] Serum: canine CAV2 standard positive serum (SN≥1:512), negative serum, ...

Embodiment 2

[0087] Example 2 Canine Passaging Reversion Test

[0088] Test Materials and Methods

[0089] Cells: Canine kidney passaged cells (MDCK), purchased from China Veterinary Drug Administration, preserved and provided by Beijing Shengtaier Technology Co., Ltd.

[0090] Virus species: Canine adenovirus type 2 virus CAV2 / 15 strain, F50 generation; preserved and provided by Beijing Shengtaier Technology Co., Ltd.

[0091] Experimental animals: The experimental dogs were 42-49-day-old healthy dogs with negative CAV2 antigen and antibody that had not been immunized with canine adenovirus type 2 vaccine.

[0092] 1. Cell passage stability test: use MDCK to passage cells, adopt the method of simultaneous inoculation, culture at 37 °C, and harvest if more than 80% cytopathic (CPE) occurs within 4 to 5 days of culture, and then continue to culture and passage with MDCK to 50%. The changes of cells at each passage (CAV2 / 15F50) were observed, and the virus titer of each passage was determi...

Embodiment 3

[0105] Example 3 Preparation of canine adenovirus type 2 live vaccine (CAV2 / 15 strain)

[0106] Preparation of attenuated antigen of canine adenovirus type 2 (CAV2 / 15 strain): CAV2 / 15 strain virus strains were inoculated into MDCK cell monolayer at a ratio of 2%, cultured at 37°C for 4 to 5 days, and freeze-thawed when the cytopathic effect reached more than 80% Harvest virus fluid.

[0107] Determination of virus titer in culture: take the cultured virus liquid as 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 Diluted and inoculated into a 96-well cell culture plate with a good monolayer of MDCK cells, each dilution inoculated into 4 wells, 0.1 mL / well, and set virus and normal cell controls at the same time, set at 37 ℃, 5% CO 2 Incubate for 4 to 5 days, observe and record CPE every day, and calculate TCID according to the Reed-Muench method 50 , virus culture virulence ≥ 10 9.0 TCID 50 / mL.

[0108] The preparation of canine adenovirus type 2 live vaccine (CAV2 / 15 strain...

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Abstract

The embodiment of the present invention relates to a canine adenovirus type 2 attenuated vaccine strain and its application. The canine adenovirus type 2 attenuated vaccine strain has a deposit number of CGMCC No.19992. The canine adenovirus type 2 (CAV2 / 15 strain) attenuated strain of the present invention is an isolated natural attenuated strain, and has the following advantages: its cultured virus price is high, and each milliliter is stable at 10 9 . 0 TCID 50 / ml; its immunization dose is low, the minimum immunization dose is 10 3 . 0 TCID 50 , using 10 5 . 0 TCID 50 It can achieve good immune effect; it can be used for dogs of various breeds and ages, and can also be used for pregnant dogs; it is safe to use, and overdose injection (injection of 20 times the immune dose) is still safe for dogs.

Description

technical field [0001] The present invention belongs to the field of biological products. The present invention relates to a canine adenovirus type 2 attenuated vaccine strain and its application. Background technique [0002] Canine adenovirus includes canine adenovirus type 1 (Canineadenovirus virus 1, CAV1) and canine adenovirus type 2 (CAV2). Canine adenovirus 2 is the main pathogen causing canine infectious laryngotracheitis (CIL). This disease is a highly contagious infectious disease that can be infected by dogs of all ages, especially puppies under 4 months old, which can cause cough in the whole litter or in the whole group, so the disease is often called "kennel cough" according to its clinical characteristics. , can lead to poor development of puppies. The main clinical symptoms are persistent high fever, cough, serous to mucous rhinorrhea, tonsillitis, laryngotracheitis, and pneumonia, and the symptoms in puppies are usually severe. Pathological changes are ma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/235A61K39/295A61K39/155A61K39/175A61K39/23A61K39/215A61P31/20A61P31/14C12R1/93
CPCC12N7/00A61K39/12A61P31/20A61P31/14C12N2710/10321C12N2710/10334C12N2750/14334C12N2760/18034C12N2760/18434C12N2770/20034A61K2039/5254A61K2039/552A61K2039/70
Inventor 金扩世薛兴华韩俊国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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