Biological specificity of 2019 novel coronavirus 614D and 614G pseudovirus
10.3760/cma.j.cn112866-20220223-00038
- VernacularTitle:新型冠状病毒614D和614G假病毒生物学特性研究
- Author:
Fenlian MA
1
;
Xiaoyi LUO
;
Chao WANG
;
Jingdong SONG
;
Zhiping XIE
;
Shanshan CONG
;
Yiman HUANG
;
Lishu ZHENG
Author Information
1. 中国疾病预防控制中心病毒病预防控制所 国家卫生健康委员会医学病毒与病毒病重点实验室，北京 100052
- Keywords:
2019 novel coronavirus;
Pseudovirus;
Neutralizing activity
- From:
Chinese Journal of Experimental and Clinical Virology
2022;36(4):397-401
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct 2019 novel coronavirus (2019-nCoV) 614D and 614G pseudovirus by HIV lentivirus packaging system and explore their biological specificity.Methods:The recombinant expression plasmids pCDNA3.1-614D and pCDNA3.1-614G were transiently cotransfected with psPAX2 and pLenti CMV Puro LUC into 293T cells respectively. After 72 hours, the supernatant was collected and ultracentrifuged with 20% sucrose cushion. The titer, morphology, protein expression and neutralizing activity of pseudovirus were determined.Results:S protein specific fluorescence was detected by indirect immunofluorescence test, Western blot analysis showed S protein was expressed, and the spike of pseudovirus was observed under transmission electron microscope. The titers of pseudovirus 614D and 614G were 1.12×10 4 and 2.52×10 4 TCID 50/ml, respectively. The pseudovirus 614D and 614G could be neutralized by S rabbit polyclonal antibody, indicating that the pseudovirus has high specificity. Conclusions:In this study, 2019-nCoV 614D and 614G pseudovirus was successfully constructed, which laid the foundation for the establishment of in vitro neutralizing antibody detection platform based on pseudovirus.
