二代测序在Omicron株新型冠状病毒灭活疫苗毒种遗传稳定性研究中的应用  

Application of next generation sequencing in the study of virus genetic stability of Omicron strain inactivated vaccine

作　　者：杨洁 郭靖 庞德钦 杨安纳 杨东升 吴杰 廖文博 周以斯 孟胜利 王泽鋆 申硕 Yang Jie;Guo Jing;Pang Deqin;Yang Anna;Yang Dongsheng;Wu Jie;Liao Wenbo;Zhou Yisi;Meng Shengli;Wang Zejun;Shen Shuo(Viral Vaccine Research Laboratory,Wuhan Institution of Biological Product Co.,Ltd.,National Engineering Technology Research Center of Combination Vaccines,Hubei Provincal Vaccine Technology Innovation Center Wuhan,Wuhan 430207,China)

机构地区：[1]武汉生物制品研究所有限责任公司病毒性疫苗研究一室,国家联合疫苗工程技术研究中心,湖北省疫苗技术创新中心,武汉430207

出　　处：《中华实验和临床病毒学杂志》2022年第5期527-534,共8页Chinese Journal of Experimental and Clinical Virology

基　　金：湖北省科技重大专项(2021ACB005);国家重点研发计划(2020YFC0842100)。

摘　　要：目的分析并监测Omicron株新型冠状病毒灭活疫苗毒种遗传稳定性。方法毒种通过不同的方式,即未经过空斑纯化的毒种和空斑纯化的毒种,在细胞上进行连续传代后,收获细胞培养上清提取病毒核酸,利用二代测序进行病毒宏转录组分析,比较不同条件下Omicron株新型冠状病毒灭活疫苗毒种的全基因组突变位点及突变频率和插入/缺失的差异。结果空斑纯化毒种连续传代后,ORF1ab和S基因序列上的高于5%的突变位点明显少于未经空斑纯化的毒种,且在纯化毒种全基因组中未检测到插入/缺失突变。结论通过二代测序技术,分析不同路径的毒种连续传代样本的全基因组序列,结果表明毒种经过空斑纯化后的遗传稳定性优于未纯化毒种,为灭活疫苗研发毒种的选育提供了科学依据。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.

关 键 词：灭活疫苗 毒种 二代测序技术 遗传稳定性 

分 类 号：R392[医药卫生—免疫学]