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Novel coronavirus and culture method thereof, and novel coronavirus inactivated vaccine

A technology of coronavirus and culture method, which is applied in the field of vaccines to achieve the effect of fast value-added, high culture titer and good passage stability

Active Publication Date: 2022-04-15
广东省疾病预防控制中心 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a new infectious disease, the new coronavirus has not yet produced a new coronavirus inactivated vaccine for human use

Method used

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  • Novel coronavirus and culture method thereof, and novel coronavirus inactivated vaccine
  • Novel coronavirus and culture method thereof, and novel coronavirus inactivated vaccine
  • Novel coronavirus and culture method thereof, and novel coronavirus inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Isolation, cultivation and identification of embodiment 1 K-T13 strain

[0070] 1. Sample processing

[0071] The K-T13 strain claimed in the present invention is isolated from samples of patients infected with the new coronavirus.

[0072] In a biosafety level 3 laboratory, the specimens are handled accordingly:

[0073] Add appropriate amount of MEM to the sputum bottle of the sputum sample submitted for inspection by a patient infected with the new coronavirus, vortex for 2 to 3 minutes to fully homogenize the sputum, and place it at room temperature. If no antibiotics were added, directly add double antibodies to a final concentration of 1000U / mL, and transfer to cryopreservation tubes.

[0074] Second, the configuration of the experimental solution

[0075]

[0076] Remarks: If the MEM medium contains glutamine, it can no longer be added; if the MEM medium does not contain glutamine, it is necessary to add a 1% (V / V) concentration of 200mM glutamine solution....

Embodiment 2

[0103] Embodiment 2 New coronavirus inactivation

[0104] (1) Virus harvest solution (37°C when harvested), equilibrate to 2-8°C before adding inactivator;

[0105] (2) β-propiolactone (stored at -20°C), put it on ice during use to ensure the temperature is below 2-8°C, and quickly return it to -20°C after use;

[0106] (3) Add the inactivator β-propiolactone into the virus harvesting solution (2-8°C) in different proportions, shake it evenly, and transfer it to a new storage bottle with a sterile pipette;

[0107] (4) Set up a negative control, add the inactivator (β-propiolactone) into the cell maintenance solution according to different proportions, shake it evenly, and transfer it to a new storage tube with a sterile pipette;

[0108] (5) Place the sample after adding the inactivator at 2-8°C for inactivation, and shake it once at a certain interval, and the frequency is higher at the beginning;

[0109] (6) After an appropriate time of inactivation, perform a 37°C water...

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Abstract

The present invention relates to the field of vaccines, in particular, to a novel coronavirus, a cultivation method thereof, and an inactivated vaccine of the novel coronavirus. The novel coronavirus was deposited in the China Center for Type Culture Collection on July 28, 2020, with the preservation number: CCTCC NO:V202039, and the strain name is 2019‑nCoV / K‑T13. The present invention provides a strain of novel coronavirus, which has high culture titer, fast multiplication, and good passage stability. Even if the strain has been passed on for more than 18 generations, the sequence of the antigenic determinant (RBD) region of the strain is still not obvious. Therefore, it is suitable for large-scale cultivation of viruses and is suitable for use as vaccine strains. The present invention also determines the optimal inactivation condition of this strain, and obtains the novel coronavirus inactivated vaccine for the first time.

Description

technical field [0001] The present invention relates to the field of vaccines, in particular, to a novel coronavirus, a cultivation method thereof, and an inactivated vaccine of the novel coronavirus. Background technique [0002] Severe acute respiratory syndrome coronavirus 2, or translated as severe acute respiratory syndrome coronavirus 2, severe acute respiratory syndrome coronavirus 2 (Severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) is a A positive-sense, single-stranded RNA virus of the membrane, belonging to the Coronaviridae Betacoronavirus genus severe acute respiratory syndrome-associated coronavirus species. Its gene sequence and SARS virus and MERS virus belong to the same lineage but different clades. It is the seventh known coronavirus that can infect humans. The hosts of the virus include mammals and birds, which caused the outbreak of coronavirus disease 2019 (COVID-19) in late 2019. The SARS-CoV-2 virus can invade the human body through the h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N7/06A61K39/215A61P31/14C12R1/93
CPCC12N7/00A61K39/12A61P31/14C12N2770/20021C12N2770/20051C12N2770/20063C12N2770/20034A61K2039/5252
Inventor 袁润余柯昌文周平平柯碧霞安文琪马小伟张海燕黎薇武婕
Owner 广东省疾病预防控制中心
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