H3N2 subtype canine influenza virus inactivated vaccine, and preparation method and application thereof

A canine influenza virus, H3N2 technology, applied in biochemical equipment and methods, antiviral agents, viruses/phages, etc.

Inactive Publication Date: 2014-07-23
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no reports of H3N8 subtype equine influenza virus infecting dogs in China

Method used

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  • H3N2 subtype canine influenza virus inactivated vaccine, and preparation method and application thereof
  • H3N2 subtype canine influenza virus inactivated vaccine, and preparation method and application thereof
  • H3N2 subtype canine influenza virus inactivated vaccine, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Isolation and identification of canine influenza virus A / Canine / Heilongjiang / L1 / 2013 (H3N2) strain of the present invention

[0027] Materials and Methods

[0028] 1. Disease materials and chicken embryos

[0029] The disease materials were collected from the lung tissue of dogs with respiratory symptoms in a pet hospital in Heilongjiang Province; SPF chicken embryos aged 9-11 days were provided by the Laboratory Animal Center of Harbin Veterinary Research Institute.

[0030] 2. Virus proliferation and HA test

[0031] After the lung tissue was ground, centrifuge at 5000rpm for 10min, take the supernatant, add 1000U double antibody, inoculate it in the allantoic cavity of 9-11-day-old SPF chicken embryos, 0.2ml / piece, culture at 37°C for 72h, overnight at 4°C, and collect the chicken embryos aseptically. Allantoic fluid, after aliquoting, stored at -70°C for later use. And determine its HA titer.

[0032] 3. Molecular biology reagents

[0033] RNA extra...

Embodiment 2

[0054] Example 2 Preparation of H3N2 subtype canine influenza virus inactivated vaccine of the present invention and evaluation of its immune efficacy

[0055] 1 Materials and methods

[0056] 1.1 Virus proliferation, inactivation and emulsification

[0057] The CIV-HLJ strain was diluted 1000 times, inoculated with 9-11-day-old SPF chicken embryos, and the chicken embryo allantoic fluid was collected at 72 h, centrifuged at 6000 rmp for 20 min, and the supernatant was taken.

[0058] 1.2 Virus inactivation and emulsification

[0059] The chicken embryo allantoic fluid was added with β-propiolactone with a final concentration of 0.2% (mass percentage concentration), and inactivated for 48 hours. Montanide PET GEL A (GEL A for short) with a final concentration of 8% (mass percentage concentration) was added, and the mixture was emulsified for 10 minutes at a shear rate of 200 rpm using a paddle stirrer.

[0060] 1.3 Analysis of Canine Immune Efficacy

[0061] Eight 10-week-...

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Abstract

The invention discloses an H3N2 subtype canine influenza virus inactivated vaccine, and preparation method and application thereof, and also discloses an H3N2 subtype canine influenza virus strain used for preparing the inactivated vaccine, and the preparation method of the inactivated vaccine. The H3N2 subtype canine influenza virus strain is named as a CIV-HLJ strain, is preserved in China General Microbiological Culture Collection Center, and has a preservation number of CGMCC NO.8760. Results of safety and immunogenicity evaluation tests of the canine influenza inactivated vaccine prepared in the invention show that the vaccine is safe to canines, and can induce the canine to to generate an ideal immune protection effect. The vaccine provides an effective means for controlling H3N2 subtype canine influenza diseases.

Description

technical field [0001] The present invention relates to an inactivated canine influenza vaccine, a preparation method thereof and a vaccine prepared therefrom. It belongs to the technical field of biological products. Background technique [0002] Canine influenza (CI) is an acute high-contact infectious disease of the respiratory tract in dogs caused by Canine influenza virus (CIV) belonging to the family Orthomyxoviridae and influenza A virus of the genus Influenza virus. It is fulminant and spreads very rapidly and widely. In 2005, CI was reported for the first time in China. The results of an epidemiological survey of CI in Guangdong by some researchers showed that the average positive rate of antibodies was 7.92% (ELISA) and 6.25% (HI). [0003] The current circulating CIV mainly includes H3N8 subtype canine influenza virus and H3N2 subtype canine influenza virus. There is no report of H3N8 subtype equine influenza virus infecting dogs in China. Since 2006, there ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/145A61P31/16
Inventor 刘大飞刘春国程景张洪英曲连东
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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