1.Application of next generation sequencing in the study of virus genetic stability of Omicron strain inactivated vaccine
Jie YANG ; Jing GUO ; Deqin PANG ; Anna YANG ; Dongsheng YANG ; Jie WU ; Wenbo LIAO ; Yisi ZHOU ; Shengli MENG ; Zejun WANG ; Shuo SHEN
Chinese Journal of Experimental and Clinical Virology 2022;36(5):527-534
Objective:To analyze and monitor the genetic stability of Omicron strain inactivated vaccine.Methods:The virus seeds of Omicron strain for inactivated vaccine through different routes, that was with plaque purification or not, were continuously passaged on cells, and then the supernatant of cell culture was harvested to extract virus nucleic acid. The next generation sequencing was used to analyze virus transcriptome, and the differences of mutation sites, mutation frequencies and insertions/deletions in the whole genome of Omicron virus under different conditions were compared.Results:After continuous passage, more than 5% mutation sites in ORF1ab and S gene sequences were significantly less in the plaque-purified seed than those of the virus without plaque purification, and no insertion/deletion mutations were detected in the whole genome of the purified virus.Conclusions:The nucleic acid sequences of virus with different routes were analyzed by next generation sequencing. The result showed that the genetic stability of virus after plaque purification was better than that of unpurified virus strains, which provides a scientific basis for virus seed selection in the development of inactivated vaccine.
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