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CPV (canine parvovirus)-HuN1703 strain and application thereof

A technology of canine parvovirus, cpv-hun1703, applied in the direction of antiviral agents, virus antigen components, viruses/bacteriophages, etc., can solve gene variation, can not fully meet the needs of disease prevention and control and market consumption, inactivated vaccines and attenuated vaccines High protection level, high antibody production level, and good immunogenicity can be achieved due to problems such as decline in immune effect

Active Publication Date: 2020-02-28
HENGYANG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as pointed out in the patent application CN201611031801.X, the CPV isolates in my country also have genetic mutations, and the results of natural host drift through the virus are very serious, because the virus can pass through innate immunity and non-adaptive The host population is widespread
Due to the rapid mutation rate, the immune effects of clinically used inactivated vaccines and attenuated vaccines have declined to varying degrees, and the existing vaccines cannot fully meet the needs of disease prevention and control and market consumption

Method used

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  • CPV (canine parvovirus)-HuN1703 strain and application thereof
  • CPV (canine parvovirus)-HuN1703 strain and application thereof
  • CPV (canine parvovirus)-HuN1703 strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Isolation and identification of canine parvovirus CPV-HuN1703 strain

[0032] 1.1 Source and collection of disease materials: collected from severely ill dogs admitted to a pet hospital in Hengyang City, Hunan Province, and disease materials after death.

[0033] It was clinically found that the dog was depressed, reluctant to eat, severe diarrhea with vomiting, red blood clots in the stool, and intestinal mucosal shedding. The existing medications can hardly prevent the deterioration of the condition. The stool was taken on-site and tested for CPV antigen with colloidal gold test strips. The test result was positive.

[0034] After the sick and dead dogs were disinfected, tissue disease materials such as the dog's heart, liver, spleen, lymph nodes, small intestine and intestinal contents were collected and stored at -70℃.

[0035] Other virus sources:

[0036] CPV standard strain (CVCC AV298), from China Veterinary Drug Administration (HA titer is 1:1024).

[0037] CP...

Embodiment 2

[0057] Example 2: Canine parvovirus CPV-HuN1703 strain is used for vaccine preparation

[0058] (1) Preparation of vaccine

[0059] An inactivated canine parvovirus vaccine prepared by the following method:

[0060] Step a, culture the cat kidney passage cells F81 according to the conventional method: the growth medium is MEM containing 5% fetal bovine serum (containing penicillin sodium and streptomycin sulfate 100 μg / mL each), at 37°C, 5% CO 2 Culture and subculture in an incubator. When the cells grow to 80% confluence, wash them with PBS once and then change to serum-free medium;

[0061] Step b, take the content as 5×10 4 TCID 50 / ml canine parvovirus CPV-HuN1703 strain was inoculated into the well-growing F81 cells prepared in step a at a 5% volume percentage, adsorbed at 37°C for 1 hour, discarded the adsorption solution, and added MEM containing 10 μg / mL trypsin Continue to cultivate as a maintenance fluid;

[0062] Step c: After inoculation, observe 3 times a day, record the c...

Embodiment 3

[0071] Example 3: Vaccine efficacy test

[0072] 3.1 Materials and methods

[0073] As a control, the CPV standard strain and the CPV-NY130615 strain were used according to the preparation method of the inactivated vaccine in Example 2 to prepare a batch of products. Chongbiwei® Youmiankang is selected as a commercial vaccine (mainly immune to canine fever, canine smallness, hepatitis and canine parainfluenza, batch number: A505A01)

[0074] Twenty-five 30-40-day-old hybrid puppies that were negative by pathogen detection and antibody detection were randomly divided into 5 groups with 5 in each group, named as control group, CPV-HuN1703 group, CPV standard group, CPV- NY130615 group and commercial vaccine group. The control group was injected with 50% ISA206 adjuvant (50% ISA206+50% saline), and the CPV-HuN1703 group, CPV standard group, and CPV-NY130615 group were injected with the 20180601 batch prepared in Example 2 The secondary vaccine is a vaccine prepared based on the CPV st...

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Abstract

The invention relates to a CPV (canine parvovirus)-HuN1703 strain and an application thereof. The blood congeal titer of the strain is equivalent to that of an existing separation toxic strain, the proliferation ability of the strain is superior to that of a CPV standard strain and a CPV toxic strain previously separated by an inventor, and the strain is a good inactivated vaccine strain. An inactivated vaccine prepared after inactivation can effectively stimulate an inoculated animal to generate an IgG antibody with effective level to cope with infection caused by different CPV toxic strains,antibody generation level is high, and animal protection capability is high.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a canine parvovirus CPV-HuN1703 strain and its application. Background technique [0002] In recent years, with the continuous improvement of people’s living standards, pets have become an indispensable part of people’s lives. While the rapidly expanding pet industry has brought vitality to social development, there are also many problems to be solved, such as canine distemper. Infectious diseases such as canine parvovirus disease and canine coronavirus disease are widespread worldwide and spread rapidly, causing great harm to the dog industry. Among them, canine parvovirus disease is particularly serious, with a fatality rate of 50% to 90%, which has seriously endangered the development of the dog industry. [0003] Canine parvovirus (CPV) is a highly contact acute and severe infectious disease characterized by vomiting, diarrhea, leukopenia, and hemorrhagic enteritis caused by...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/23A61P31/20C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552A61P31/20C12N7/00
Inventor 唐青海杨海王芳宇杨灿何丽芳唐娇玉曹丽敏易诚唐斯萍
Owner HENGYANG NORMAL UNIV
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