Application of H3N2 canine influenza virus CGD1

A technology of canine influenza virus and H3N2, which is applied in the direction of antiviral agents, medical preparations containing active ingredients, antibody medical ingredients, etc., can solve the problems of large differences in antigenicity of canine influenza virus, shorten the detoxification period and improve prevention and therapeutic effects, severity-reducing effects

Active Publication Date: 2013-07-31
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the deficiency that the antigenicity of cani

Method used

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  • Application of H3N2 canine influenza virus CGD1
  • Application of H3N2 canine influenza virus CGD1
  • Application of H3N2 canine influenza virus CGD1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation of H3N2 canine influenza virus.

[0030] S1. Sample collection and processing:

[0031] Use sterile cotton swabs to collect nasopharyngeal secretions in dogs. When collecting nasal swabs, try to wipe the secretions from the nose or insert the swab deep into the nasal cavity, and pull it out after rotating for several weeks; when collecting throat swabs, try to Wipe the secretions of the throat more. The collected swabs were placed in 2mL EP tubes, soaked in sterilized saline containing penicillin and streptomycin, and sent to the laboratory for testing or stored at -20°C.

[0032] S2. Chicken embryo inoculation:

[0033] Take out the nasopharyngeal swab after repeatedly squeezing the tube wall, and fully shake the liquid in the tube. Place at 4°C and wait for natural precipitation for 5-10 minutes, take the supernatant and add double penicillin and streptomycin with a final concentration of 1 000 μg / mL, act at 4°C for 1 hour, inoculat...

Embodiment 2

[0052] Example 2 Breeding and selection of H3N2 canine influenza vaccine strains.

[0053] Breeding of S1.H3N2 Canine Influenza Vaccine Strains

[0054] A / canine / Guangdong / 01 / 2006(H3N2), A / canine / Guangdong / 02 / 2006(H3N2), A / canine / Guangdong / 01 / 2007(H3N2) isolated from 2006~2007 are referred to as Three strains of H3N2 canine influenza virus, CGD1, CGD2, and CGD3, were used as vaccine candidate strains. 3 strains of vaccine candidate strain virus allantoic fluid are first made 10 -1 ~10 -3 Inoculate 9-day-11-day-old SPF chicken embryos through the allantoic cavity, the inoculation volume is 0.2ml / embryo, inoculate 5 eggs for each dilution, incubate at 37°C for 72-96h, discard dead embryos within 24 hours, Observed every 12 h, and stored the dead embryos in a refrigerator at 4 °C. After 96 hours, cool the surviving chicken embryos at 4°C overnight, collect the allantoic fluid of the chicken embryos aseptically, and measure the hemagglutination units (HAU) with 1% chic...

Embodiment 3

[0087] Example 3 Purity test of H3N2 canine influenza vaccine strains.

[0088] S1. Bacterial test: According to the relevant methods in the "Quality Standards for Veterinary Biological Products of the People's Republic of China 2001 Edition", take the newly harvested virus allantoic fluid and inoculate them with thioglycolate medium (T.G) and glucose-peptone medium respectively. (G.P), modified Frey's medium, cultured at 37°C for 72 hours, and observed whether there was microbial growth. (Detect bacteria, mold and mycoplasma)

[0089] Inspection medium configuration method:

[0090] Thioglycollate medium (T.G): tryptone 15.0g; agar (powder) 0.5~0.7g; yeast extract powder 5.0g; sodium chloride 2.5g; glucose 5.0g; 0.2% methylene blue solution (or 1 % resazurin solution 1.0ml) 0.5ml; sodium thioglycolate 0.5g; L-cysteine ​​hydrochloride 0.5g; water for injection was added to 1000ml. Mix the above ingredients, heat to dissolve, and adjust the pH to 7.0~7.2 with sodium ...

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Abstract

The invention belongs to the technical field of preparation of virus vaccine, and concretely discloses an application of an H3N2 canine influenza virus CGD1. According to the invention, three self-isolated, identified and preserved strains of H3N2 canine influenza virus CGD1, CGD2 and CGD3 are used to inoculate chick embryo for subcultring, and the CGD1 is found to be good in genetic stability. Therefore, the CGD1 is selected for plaque purification to breed an H3N2 canine influenza virus vaccine strain CIVGDYM1, and the vaccine strain has been preserved in China general microbiological culture collection center, with an accession number being CGMCCNO: 7218. By using the H3N2 canine influenza virus vaccine strain CIVGDYM1 to inoculate the chick embryo for virus breeding, and through gaining allantoic fluid of the chick embryo, inactivating, preparing vaccine and other steps, the safe and effective H3N2 canine influenza virus inactivated vaccine can be prepared.

Description

technical field [0001] The invention relates to the technical field of virus vaccine preparation, and more specifically relates to the application of a H3N2 canine influenza virus strain CGD1. Background technique [0002] Influenza A virus is an important infectious disease that threatens human health. Influenza viruses have strict host specificity, and even the spread of the same virus on different hosts is restricted by the host community. Traditionally, dogs and cats are not susceptible to influenza virus, but influenza virus infection has been reported in these two animals in recent years. Experimental and natural infections have demonstrated that H5N1 subtype avian influenza can infect dogs. As early as 2004, the United States reported for the first time that the H3N8 subtype of canine influenza virus caused a major outbreak of canine influenza. Through sequence analysis, it was found that this subtype of canine influenza virus was derived from equine influenza virus...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61P31/16
Inventor 李守军赵明喜李华涛贾坤孙凌霜远立国王衡
Owner SOUTH CHINA AGRI UNIV
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