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Human parainfluenza virus type 3 (HPIV-3) wild strain and application thereof

A technology of parainfluenza and wild strains, applied in the direction of viruses, viral peptides, antiviral agents, etc., can solve the problems of drugs and vaccines without specific effects, and achieve the effect of preventing infection

Active Publication Date: 2019-12-31
LANZHOU INST OF BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the prevention and treatment of HPIV-3 infection, there are currently no specific drugs and vaccines, and the WHO lists HPIV-3 vaccines as a priority vaccine for future development

Method used

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  • Human parainfluenza virus type 3 (HPIV-3) wild strain and application thereof
  • Human parainfluenza virus type 3 (HPIV-3) wild strain and application thereof
  • Human parainfluenza virus type 3 (HPIV-3) wild strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Isolation and purification of HPIV-3 wild strain

[0045] 1. Isolation of HPIV-3 wild strain

[0046] Candidate strains must have complete records, history, and sources. Determine the biological characteristics of the candidate virus strains. The attenuated live vaccines and subunit vaccines prepared by the candidate virus strains should have high virus yields, strong ability to induce immune protection, broad cross-protection spectrum, and stable biological characteristics. Virus strains with broad epidemic potential at present and in the future selected by molecular epidemiology and molecular epidemiology.

[0047] The source of the virus: the lower respiratory tract secretions of children hospitalized with lower respiratory tract infection in Maternal and Child Health Hospital of Gansu Province.

[0048]Isolation of virus strains: Add 0.5ml of the obtained secretion into 3ml of DMEM (Dulbecco's Modified Eagle Medium, containing various amino acids and glu...

Embodiment 2

[0067] Example 2 Determination of virus titer

[0068] Determination of virus titer by plaque assay

[0069] HPIV3LZ1728C19 virus was diluted 10x serially, inoculated with MA104 cells in 6-well plates, and placed at 37°C, 5% CO 2 Incubate in an incubator for 2 hours, suck out the supernatant, add 3ml of 0.8% low melting point agarose, after solidification at room temperature, place it upside down at 37°C, 5% CO 2 Cultivate in an incubator for 72 hours, add 3ml of 1% low-melting point agarose containing neutral red, and solidify at room temperature at 37°C, 5% CO 2 Incubate overnight, observe plaques, and take the highest dilution of plaques as the virus titer.

[0070] The highest virus titer obtained by plaque assay was 10 9 pfu / ml, it can be seen that the HPIV3LZ1728C19 virus can obtain a higher virus titer through cell culture.

Embodiment 3

[0071] A large amount of preparation and purification of embodiment 3 HPIV-3 virus culture fluid

[0072] 1. Preparation of HPIV-3 virus culture solution.

[0073] Inoculate the HPIV3LZ1728C19 virus strain, which was cloned and purified to remove specific peripheral factors, into WI-38 cells at 0.05 MOI, at 37°C, 5% CO 2 Culture in an incubator for 7 days, collect the culture supernatant, detect the virus titer, and freeze at -80°C.

[0074] 2. The HPIV-3 virus was purified by ultracentrifugation.

[0075] The HPIV3LZ1728C19 virus culture solution was centrifuged at 8000rpm at 4°C for 30min to precipitate cell debris and collect the supernatant. Take 30ml of the supernatant and add it to a 50ml special centrifuge tube, gently push in 15ml of 50% sucrose from the bottom with an 8cm long needle, pay attention to keep the interface clear, put it into an ultracentrifuge (model: Hitachi CP 70MX) for 4 Centrifuge at 35,000 rpm for 4 hours, discard the supernatant; resuspend the p...

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Abstract

The invention relates to a human parainfluenza virus type 3 (HPIV-3) wild strain and an application thereof. The preservation number of the wild strain is CCTCC NO:V201911, the wild strain has HN protein and F protein of an HPIV-3 virus, and the wild strain can be applied to an HPIV-3 virus infected mouse model and an HPIV-3 neutralizing antibody detection experiment, and can be applied to preparation of vaccines for preventing parainfluenza type 3. The human parainfluenza virus type 3 wild strain disclosed by the invention is a cloned purified high-titer HPIV3LZ1728C19 virus strain free fromspecific exogenous factor plaque formation, and a human parainfluenza virus type 3 HN protein subunit vaccine prepared from the virus can effectively prevent parainfluenza virus type 3 infection.

Description

technical field [0001] The invention belongs to the field of type III parainfluenza virus, in particular to a type III parainfluenza virus wild strain and application thereof. Background technique [0002] Human parainfluenzavirus type 3 (HPIV-3) is a pathogen that causes severe lower respiratory tract diseases in infants and young children after respiratory syncytial virus. It mainly causes pneumonia and bronchitis in infants under one year old. In addition, HPIV-3 is also an important cause of lower respiratory tract disease in elderly patients with chronic diseases and immunocompromised adults. Outbreaks of HPIV-3 occur every year, mainly in spring and summer. In the United States, the number of hospitalizations caused by HPIV-3 infection is estimated to be up to 30,000 people each year, resulting in a large disease burden. In developing countries, although there is no specific data on the disease burden caused by HPIV-3, it is determined by HPIV- 3 The lower respirato...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C07K14/115C07K16/10C07K1/22G01N33/569A61K39/155A61P31/14C12R1/93
CPCC12N7/00C07K14/005C07K16/1027G01N33/56983A61K39/12A61P31/14C12N2760/18621C12N2760/18622G01N2333/115C12N2760/18634
Inventor 白慕群关文竹火文马超傅生芳李雄雄陈汉泉寇桂英包红
Owner LANZHOU INST OF BIOLOGICAL PROD
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