1. Fujian Center for Disease Control and Prevention, Fujian Provincial Key Laboratory for Zoonoses Research, Fuzhou 350001, China; 2. School of Public Health, Fujian Medical University, Fuzhou 350108, China
Abstract:This study aimed to explore the characteristics of 2019-nCoV proliferation on Vero-E6 cells and to determine the necessary experimental conditions for viral isolation and culture, antiviral drug research and vaccine development. The TCID50 of the 2019-nCoV strain isolated from Fujian province was determined, and virus (1.74×10-4TCID50/cell) was inoculated into the Vero-E6 cell-line. The supernatants of infected cells were collected every 24 hours for subsequent viral titration and determination of the number of genomic copies at 0-6 days post-infection. The effects of different experimental conditions on the TCID50, such as freeze-thaw of viral stock, cell concentrations for viral inoculation and washing after viral adsorption, were explored. The results showed that the 2019-nCoV virus replicated rapidly on Vero-E6 cells in the first 48 hours, and the average replication period was approximately 3.29 h. The nucleic acid concentration and viral titer peaked at approximately 48 hours and thereafter gradually declined, but the nucleic acid concentration remained high. Freeze-thaw of viral stock, cell concentration and washing of the virus-adsorbed cells did not significantly affect the viral TCID50.
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