A Strain of Feline Parvovirus and Its Application

A feline parvovirus and strain technology, applied in the direction of viruses, antiviral agents, viruses/bacteriophages, etc., can solve the problems of limited purchase channels, high prices, and no FPV vaccine approved for use, and achieve good safety and good protection force effect

Active Publication Date: 2022-05-24
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the imported cat triple inactivated vaccine (FPV, FCV, FHV) is basically used in the market, but imported vaccines have disadvantages such as high price, limited purchase channels, and prone to fake and shoddy products.
[0006] After data inquiry and information check, no domestic FPV vaccine has been approved for use

Method used

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  • A Strain of Feline Parvovirus and Its Application
  • A Strain of Feline Parvovirus and Its Application
  • A Strain of Feline Parvovirus and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Isolation and identification of FPV WH1

[0031] 1. Experimental method

[0032] 1. Virus isolation

[0033] (1) The patient materials diagnosed with feline parvovirus were collected from a pet hospital in Wuhan, China, most of which were feces. Dilute the feces with 10 times the volume of sterile 4.5g / L DMEM, mix well (operate on ice), 4°C, 9000rpm, 20min. The supernatant was collected by centrifugation, filtered through a 0.22 μm filter, and dispensed into sterile EP tubes as the virus to be separated, and stored at -80°C.

[0034] (2) Cultivate the susceptible cell CRFK to a cell density of 50%-60% in a T25 cell culture flask, inoculate the diseased material treated in the previous step into the susceptible cell CRFK at a ratio of 1:10, and place it at 37°C for culture Incubator, adsorbed for 2h, then changed to 2% DMEM maintenance solution, put it into 37 ℃, 5% CO 2 Culture in the incubator for 3-5 days and observe whether any lesions appear.

[0035]...

Embodiment 2

[0049] Example 2 Plaque purification of FPV WH1

[0050] 1. Experimental method

[0051] (1) After evenly digesting CRFK, adjust the cell concentration and inoculate a 6-well plate at an appropriate concentration. When the cell density is 50%-60%, prepare a virus solution and dilute it tenfold to obtain five concentrations of virus solution. 100 μL of virus solution was added dropwise to each well, adsorbed at 37°C for 2 hours, and the plate was shaken every 15 min to fully absorb the virus solution. After 2 hours, the virus solution was aspirated and washed twice with PBS.

[0052] (2) Prepare a 2% low-melting point agarose solution, melt at 72°C, and place it in a 37°C water bath to keep warm for later use. 2×DMEM is also preheated in a 37°C water bath.

[0053] (3) Mix the above 2% low agarose solution with 2×DMEM maintenance solution (2% serum, 1% double antibody) in a ratio of 1:1, add 2 mL / well to each well, and add 2 mL / well to a 6-well plate. Put it in the refrigerat...

Embodiment 3

[0057] Example 3 Determination of FPV WH1 Growth Kinetics

[0058] 1. Experimental method

[0059] (1) Inoculate the cells in a 24-well plate, and when the cell density reaches about 60%, infect the cells with 0.1MOI virus, and at the same time, replace it with 2% DEME maintenance solution (2% serum, 1% double antibody), and place it in 37 ℃ in an incubator.

[0060] (2) Collect cell supernatant samples at 12, 24, 36, 48, 60, 72, and 84 hours after inoculation, and use CRFK cells to detect viral TCID 50 .

[0061] (3) Subculture the cells in a 96-well plate, and when the cell density reaches about 50%, dilute the virus by 10 times with cell maintenance solution, add 100 μL of virus solution to each well, and repeat 8 times for each dilution. , placed in an incubator.

[0062] (4) Take out after 60 hours, use the indirect immunofluorescence method to measure the result of the virus titer, and use the Reed-Muench method to calculate the virus TCID 50 .

[0063] 2. Experime...

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Abstract

The invention discloses a feline parvovirus strain and application thereof, belonging to the technical field of biological products. The preservation number of the virus strain is: CCTCC NO:V202127, the preservation time is: April 27, 2021, and the preservation address is: China. Wuhan. Wuhan University. The virus strain belongs to the isolate strain in Wuhan, China, and has a high virus titer; the inactivated product of the virus can produce a good immune response in animals, has good immunogenicity, and can be used as an inactivated vaccine Stimulate the body to produce a high level of neutralizing antibodies against FPV, which has the basic potential of a vaccine. The virus strain prepared into a vaccine can be used in the prevention and treatment of feline panleukopenia, so as to provide an important vaccine virus source for the prevention and control of feline panleukopenia in my country, and realize the effective prevention and control of feline parvovirus.

Description

technical field [0001] The invention relates to the technical field of biological products, and relates to a feline parvovirus strain and its application. Background technique [0002] Feline panleukopenia, also known as feline distemper, feline distemper, and feline infectious enteritis, is a highly contagious acute infectious disease of cats. Mainly occurs in young cats (2 months old - 4 months old), the prevalence and mortality are relatively high, the mortality rate is as high as 50% -70%, known as the kitten killer. Adult cats have a lower risk of disease, and most of them have latent infection or subclinical infection. In addition to infecting domestic cats, it can also infect other wild cats, such as tigers, bears, leopards, etc. The typical clinical symptoms of sick cats are anorexia, lethargy in the initial stage, vomiting, elevated body temperature (40℃±), diarrhea in the middle stage, blood in the stool, dehydration, leukocyte reduction below the detection line,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/23A61P31/20
CPCC12N7/00A61K39/12A61P31/20C12N2750/14021C12N2750/14034A61K2039/5252A61K2039/552A61K2039/575
Inventor 彭贵青李利沙沈洲刘紫微汪娇廖英飞
Owner HUAZHONG AGRI UNIV
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