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Novel coronavirus and culture method thereof as well as novel coronavirus inactivated vaccine
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A technology of coronavirus and culture method, applied in the field of vaccines, to achieve the effect of fast value-added, good passage stability, and high culture titer
Active Publication Date: 2020-10-13
广东省疾病预防控制中心 +2
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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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Embodiment 1
[0069] Isolation, cultivation and identification of embodiment 1 K-T13 strain
[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 sputumbottle 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 Inactivation of new coronavirus
[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 wa...
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Abstract
The invention relates to the field of vaccines, in particular to a novel coronavirus and a culture method thereof as well as a novel coronavirusinactivated vaccine. The novel coronavirus was preserved in China Center for Type Culture Collection on July 28th, 2020, with the preservation number of CCTCC NO: V202039 and the strain name of 2019-nCoV / K-T13. The invention provides a strain of the novelcoronavirus, the strain has high culture titer, fast proliferation and good passage stability, and the antigenic determinant (RBD) regional sequence of the strain is still not changed obviously evenif the strain is passed for more than 18 generations, so that the strain is suitable for large-scale culture of viruses and is suitable for being used as a vaccine strain. The optimal inactivation condition of the strain is further determined, and the novel coronavirus inactivated vaccine is obtained for the first time.
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