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Cat parvovirus strain and application thereof

A feline parvovirus and strain technology, applied in the direction of viruses, antiviral agents, virus antigen components, 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: 2021-09-03
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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  • Cat parvovirus strain and application thereof
  • Cat parvovirus strain and application thereof
  • Cat parvovirus strain and application thereof

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) Feline parvovirus-confirmed disease materials were collected from a pet hospital in Wuhan, China, and most of the disease materials were feces. The feces were diluted with 10 times the volume of sterile 4.5g / L DMEM, mixed (operated on ice), 4°C, 9000rpm, 20min. The supernatant was collected by centrifugation, filtered with a 0.22 μm filter, and distributed into sterile EP tubes, as the virus to be isolated, and stored at -80°C.

[0034] (2) Cultivate the susceptible cell CRFK in a T25 cell culture flask to a cell density of 50%-60%, inoculate the susceptible cell CRFK with the disease material treated in the previous step at a ratio of 1:10, and culture it at 37°C Box, adsorb for 2 hours, then replace with 2% DMEM maintenance solution, put in 37℃, 5% CO 2 Cultivate in an incubator for 3-5 days, and observe whether there are lesions.

[0035] (3...

Embodiment 2

[0049] Example 2 Plaque purification of FPV WH1

[0050] 1. Experimental method

[0051] (1) After digesting CRFK evenly, adjust the cell concentration, and inoculate a 6-well plate with an appropriate concentration. When the cell density is 50%-60%, prepare the virus solution and perform ten-fold dilution to obtain five concentrations of the virus solution. Add 100 μL of virus liquid to each well respectively, absorb at 37°C for 2 hours, shake the plate every 15 minutes to fully absorb the virus liquid, discard the virus liquid after 2 hours, and wash twice with PBS.

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

[0053] (3) Mix the above 2% low agarose solution with 2×DMEM maintenance solution (2% serum, 1% double antibody) according to the ratio of 1:1, add to each well, 2mL / well, put 6-well plate Put it in the refrigerator at 4°C for 10 minut...

Embodiment 3

[0057] Example 3 Determination of Growth Kinetics of FPV WH1

[0058] 1. Experimental method

[0059] (1) Cells were subcultured in a 24-well plate, and when the cell density reached about 60%, the cells were infected with 0.1 MOI virus, and at the same time replaced with 2% DEME maintenance solution (2% serum, 1% double antibody), placed at 37 Cultivate in an incubator.

[0060] (2) Cell supernatant samples were collected at 12, 24, 36, 48, 60, 72, and 84 hours after inoculation, and CRFK cells were used to detect virus TCID 50 .

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

[0062] (4) Take it out after 60 hours, measure the poison price result by indirect immunofluorescence method, and calculate the virus TCID by Reed-Muench m...

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Abstract

The invention discloses a cat parvovirus strain and application thereof, and belongs to the technical field of biological products. The preservation number of the strain is CCTCC NO: V202127, the preservation time is April 27, 2021, and the preservation address is Wuhan University, Wuhan City, China. The strain belongs to an isolated strain in Wuhan of China, and has high virus titer; an inactivated product of the virus can generate a good immune response reaction in animals, has good immunogenicity, can well stimulate the body to generate a high-level neutralizing antibody for FPV as an inactivated vaccine, and has the basic potential of vaccines. The vaccine prepared from the virus strain can be used for preventing and treating cat leukopenia, so that an important vaccine virus source is provided for prevention and control of the cat leukopenia in China, and effective prevention and control of cat parvovirus are realized.

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. It mainly occurs in young cats (2-4 months old), and the morbidity and mortality are relatively high. The mortality rate is as high as 50%-70%. It is called a kitten killer. Adult cats have a lower chance of getting sick, and most of them are subclinical infections or subclinical infections. In addition to infecting domestic cats, it can also infect other wild cats, such as tigers, bears, and leopards. The typical clinical symptoms of sick cats are anorexia and listlessness in the early stage, vomiting in the middle stage, elevated body temperature (40℃±), diarrhea, and blood in the stool, dehydration, decreased white blood cells be...

Claims

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Application Information

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Patent Type & Authority Applications(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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