Inactivated vaccine for CH-JL5 strain of Feline calicirus

A CH-JL5, virus inactivation technology, applied in the direction of vaccines, viruses, antiviral agents, etc., can solve the problems of inability to eliminate, reduce the immune protection effect of vaccines, and no vaccine virus isolates, and achieve the effect of convenient operation.

Active Publication Date: 2016-12-07
CHANGCHUN SR BIOLOGICAL TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Vaccination can control the spread of the disease to a certain extent, but it cannot eliminate it; maternal antibodies can also control the disease of susceptible animals to a certain extent, but not all of them; It is a carrier of the virus, and it excretes the virus to the outside world irregularly, causing infection in healthy animals or becoming a carrier of the virus
[0004] Vaccination is the best preventive measure against the disease. Due to the variability of FCV strain antigens, it means that no vaccine can neutralize all virus isolates, thus reducing the immune protection of vaccines, and even some vaccines total failure problem

Method used

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  • Inactivated vaccine for CH-JL5 strain of Feline calicirus
  • Inactivated vaccine for CH-JL5 strain of Feline calicirus
  • Inactivated vaccine for CH-JL5 strain of Feline calicirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Isolation and Identification of Feline Calicivirus CH-JL5 Strain

[0024] 1. Isolation of feline calicivirus CH-JL5 strain

[0025] 1. In May 2013, the oral saliva of clinically suspected cats in Jilin Province was collected with oral cotton swabs;

[0026] 2. Sample processing: Add 500 μL PBS buffer solution to the cat’s oral cotton swab for 5 minutes, centrifuge at 3000 r / min for 10 minutes, discard the cotton swab, mix and separate out 250 μL for later use;

[0027] 3. Isolation and culture of virus: centrifuge the sample at 3000r / min for 15min, take the supernatant and add the same amount of DMEM nutrient solution, filter and sterilize with a 0.22μm filter, inoculate F81 monolayer cells, and place at 37°C in 5% CO 2 When cultured in an incubator, obvious lesions appeared in the second passage after blind passage, and the cytopathic changes were stable at the fifth passage, showing that the cells shrunk round and looked like grape bunches, and finally fell...

Embodiment 2

[0036] Embodiment 2 The preparation method of feline calicivirus inactivated vaccine

[0037] 1. F81 cell recovery: Take out the ampoule or cryovial from the liquid nitrogen tank, put it into a 37°C water bath quickly, and shake it from time to time to completely melt it within 1-2 minutes;

[0038] 2. F81 cell culture: Take out the cells under sterile conditions, centrifuge at 1000r / min for 5min, discard the supernatant, add 1ml 10%DMEM along the tube wall, do not blow up the cells; then add 1ml 10%DMEM, blow evenly cell. Add the cell solution to a T25 cell culture flask, add 5-6ml 10% DMEM to the cell culture flask, mix gently and store at 37°C, 5% CO 2 Cultivate in the incubator and observe twice a day;

[0039] 3. FCV virus amplification: The multiplicity of infection (MOI) was used to determine the amount of virus inoculation, and the FCV was diluted to 0.1, 0.01, and 0.001 respectively, and then inoculated into F81 cells. The virus titers of several infectious amounts ...

Embodiment 3

[0044] Embodiment 3, sterility test and safety experiment

[0045] Take a small amount of above-mentioned inactivated FCV seedlings and inoculate them in dextrose-peptone broth (GP), thioglycolate medium (TG), and casein agar (GA), and detect whether there is bacterial growth according to conventional methods. If there is no bacterial growth, add thimerosal sodium in the sterile room at a rate of 1 / 10,000 of the total amount, then divide into 10 mL per bottle, stopper, seal, label and store in a 4°C refrigerator for later use. Fifteen experimental cats aged 45-60 days were randomly divided into 3 groups, and inoculated with FPV inactivated vaccine 0.5ml, 1ml, 2ml respectively, and 5 negative control cats were not injected with the vaccine, but were raised in the same room as the injected cats, and the vaccine was injected within 15 days Clinical manifestations were observed daily.

[0046] The results showed that the prepared cell cultured inactivated vaccine had no bacterial...

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Abstract

The invention discloses a CH-JL5 strain of Feline calicirus and an inactivated vaccine for the CH-JL5 strain of the Feline calicirus. The inactivated vaccine is used in a subcutaneous injection manner, and the immunization operation is convenient. Experimental results show that the inactivated vaccine does not cause toxicity enhancement of FCV or gene integration as well as conjunctivitis, rhinitis, dental ulcer or acute death of cats, is safe and has no side effect. Experiments show that inactivated vaccine for the Feline calicirus can induce the cats to generate good humoral immunity and cellular immunity, and by carrying out a virus attack test by virtue of an FCV CH-SH strain, the immune protective rate is 96%; by carrying out the virus attack test by virtue of an FCV 2080 strain, the immune protective rate is 100%; and by carrying out the virus attack test by virtue of an FCV USDA strain, the immune protective rate is 100%.

Description

technical field [0001] The invention belongs to the field of molecular biology and immunology, and in particular relates to feline calicivirus CH-JL5 strain, feline calicivirus CH-JL5 strain inactivated vaccine and a preparation method thereof. Background technique [0002] Feline calicivirus ( Feline calicivirus , FCV), is a pathogen that can cause oral and upper respiratory diseases in cats and all cats. It is a multiple, epidemic and highly contagious pathogen. - Rhinoconjunctivitis. FCV is a kind of virus that is easy to mutate. Because of the existence of mutated strains, the virulence of different strains is different for different animal physiques, and the clinical symptoms are also different. FCV infection alone does not cause death in cats and felines, and mixed infections with bacteria, chlamydia, mycoplasma, parasites and other feline upper respiratory viruses often occur, which is one of the reasons for the increased mortality of the disease. In recent years...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/06A61K39/125A61P31/14
CPCA61K39/125A61K2039/5252A61K2039/552C12N7/00C12N2770/32021C12N2770/32034
Inventor 夏振强胡桂学王开金宏丽应瑛陈小庆
Owner CHANGCHUN SR BIOLOGICAL TECH
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