Novel coronal virus strain and medicinal use thereof
A coronavirus, a new type of technology, applied in antiviral agents, pharmaceutical formulations, viruses/bacteriophages, etc., can solve problems such as lack of support
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Embodiment 1
[0024] Throat swab specimens were taken from patients with confirmed SARS, treated with 50 micrograms / ml penicillin, 50 micrograms / ml streptomycin, and 1 microgram / ml amphotericin, and inoculated with Vero E6 cells and HEP-2 cells respectively. 2 Cultivate at 35°C, and observe the cytopathic effect (CPE) every day. After 7 days of culture, the supernatant of the diseased cells is ultracentrifuged with a sucrose gradient, and the virus particles are separated for negative staining observation with an electron microscope. The diameter of the coronavirus-like particles can be seen At 60-100nm, the virus morphology is consistent with the novel coronavirus (SARS-CoV).
Embodiment 2
[0026] Separate the Vero E6 cells infected with the virus and make the cell sheet. When the cell is pathological, take out the infected cell droplet and put it in CO 2Incubate at 35°C to allow the cells to adhere to the wall. Remove the slides, rinse with PBS, remove unattached cells, and air dry. Put it in cold acetone at -20°C for 30 minutes, and dry it in air. Drop 1:10 dilution of convalescent serum from a confirmed SARS patient three weeks later, put it into an incubator at 37°C for 30 minutes, wash with 0.01mol / L pH7.2 PBS three times, and absorb the liquid. Fluorescently labeled goat anti-human IgG antibody, purchased from CHEMICON, was added dropwise, placed in a warm box at 37°C for 30 minutes, washed three times with 0.01mol / L PBS pH7.2, and the liquid was sucked off. After indirect immunofluorescence staining, it can be seen that the edge of the infected cells is stained in a half-moon shape, and the fluorescence is obvious. Normal Vero E6 cells have no fluoresce...
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