Feline herpesvirus I type virus strain and application thereof

A feline herpes virus and virus strain technology, applied in the field of biological products, can solve problems such as inability to effectively control, antibody titer not reaching the titer against virus infection, poor immune effect, etc.

Active Publication Date: 2021-08-27
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that the neutralizing antibody qualified rate of clinical pet cats immunized with FHV-1 vaccine is only 60%, and the antibody titer cannot reach the titer of resisting virus infection. The clinical symptoms caused by -1, but its immune effect is not good, can not effectively control the current prevalence of FHV-1 in my country
This may be because the vaccine strains used in clinical practice are not the current epidemic strains in my country, and the antibody titers after vaccine immunization cannot meet the requirements

Method used

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  • Feline herpesvirus I type virus strain and application thereof
  • Feline herpesvirus I type virus strain and application thereof
  • Feline herpesvirus I type virus strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Isolation and identification of embodiment 1FHV-1 / WH / 2020 virus strain

[0029] 1. Experimental method

[0030] 1. Virus isolation

[0031] (1) The eye and nose swab samples of cats diagnosed with FHV-1 infection were collected from a pet hospital in Wuhan, China, and added with serum-free DMEM culture medium (containing 1% double antibody) to make a 1:5 suspension, centrifuged at 5000r / min for 10min, The supernatant was taken, sterilized by filtration through a 0.22 μM microporous membrane, and dispensed into sterile EP tubes as the inoculum for the isolated virus, and stored at -80°C.

[0032] (2) Cultivate FHV-1 susceptible cells CRFK to a monolayer in a 6-well culture plate, inoculate the diseased material samples treated in the previous step into CRFK according to 1 / 10 of the volume of the culture solution, and inoculate in a 37°C incubator After 1-2 hours of adsorption, the culture medium was discarded, and the CRFK cells were washed three times with PBS. Add D...

Embodiment 2

[0042] The plaque purification of embodiment 2FHV-1 / WH / 2020

[0043] 1. Experimental method

[0044] (1) After digesting CRFK evenly, adjust the cell density to 1×10 5 Cells / mL were inoculated into a 6-well plate. When the cells grew to a single layer, the virus solution was prepared, and ten-fold serial dilutions were performed to obtain five dilutions of the virus solution. Discard the culture medium in the wells, wash the wells with PBS 3 times, inoculate 800 μL of the virus solution into each well, absorb at 37°C for 2 hours, shake the plate every 15 minutes to fully absorb the virus solution, and discard the virus solution after 2 hours , washed three times with PBS.

[0045] (2) Prepare 2% low-melting point agarose solution, melt it at 72°C, put it in a water bath at 42°C to keep it warm for use, and place 2×DMEM in a water bath at 37°C for preheating.

[0046] (3) Mix the above 2% low-melting point agarose solution with 2×DMEM maintenance solution (2% serum, 1% doubl...

Embodiment 3

[0050] The determination of embodiment 3FHV-1 / WH / 2020 growth kinetics

[0051] 1. Experimental method

[0052] (1) CRFK cells were subcultured and inoculated in 24-well plates. After the cells grew to a single layer, the cells were infected with 0.01 MOI virus, and the culture medium was replaced with 2% maintenance medium, and cultured in a 37°C incubator.

[0053] (2) Supernatant samples were collected at 12, 24, 36, 48, 60, and 72 hours after inoculation, and the virus TCID was measured with CRFK cells 50 .

[0054] (3) Cells were subcultured in a 96-well plate, and when the cell density reached about 50%, the virus was diluted 10 times with the cell maintenance solution, and 100 μL of the virus solution was added to each well, and each dilution was replicated 8 times. Put into the incubator to cultivate.

[0055] (4) Observe once every 24 hours, record the virus dilution and the number of holes where cytopathic disease occurs, stop the observation after 4-5 days, record...

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Abstract

The invention discloses a feline herpesvirus I type virus strain and application thereof, and belongs to the technical field of biological products. The preservation number of the virus strain is CCTCC NO: V202126, the preservation time is April 27, 2021, and the preservation address is Wuhan University, Wuhan, China. The virus strain belongs to an isolated strain in Wuhan of China, has high virus titer of 107.65 TCID50 / mL, has strong pathogenicity to domestic cats and has the basic potential of becoming a vaccine candidate strain, a vaccine prepared from the virus strain can be used for preventing and treating feline infectious rhinotracheitis diseases, an important vaccine virus source is provided for prevention and control of the feline herpesvirus I in China, and effective prevention and control of the feline herpesvirus I are achieved.

Description

technical field [0001] The invention relates to the technical field of biological products, and relates to a type I feline herpes virus strain and an application thereof. Background technique [0002] Feline infectious rhinotracheitis, also known as feline nasal bronchitis, is a highly contagious infectious disease characterized by acute upper respiratory symptoms. Specific symptoms include: keratoconjunctivitis, upper respiratory tract infection and abortion, but sneezing, excessive Salivation, increased eye and nose secretions and other upper respiratory symptoms were the main symptoms. The disease is common in cats clinically, the incidence rate of cats is as high as 100%, and the fatality rate varies greatly among cats of different ages. Adult cats generally do not cause death, but the mortality rate of young cats can reach 50%. Sick animals can carry toxins and detoxify all their lives, and they can be repeatedly infected under certain conditions. Crandell and Maurer ...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N15/38A61K39/245A61P31/22C12R1/93
CPCC12N7/00C07K14/005A61K39/12A61P31/22C12N2710/16721C12N2710/16722C12N2710/16734A61K2039/5252A61K2039/5254A61K2039/53A61K2039/552
Inventor 彭贵青杨梦芳沈洲刘紫微汪娇廖英飞
Owner HUAZHONG AGRI UNIV
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