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为了筛选获得可用于人冠状病毒OC43(Human coronavirus OC43,HCoV-OC43)分离与研究的细胞系,本研究利用表达海肾荧光素酶基因的重组HCoV-OC43(rOC43-ns2DelRluc)对28种常见真核细胞系进行了易感性高效筛选研究,进一步利用亲本病毒HCoV-OC43-WT进行了细胞病变效应的观察,并采用荧光定量PCR进行复制动力学分析。实验结果显示:18种细胞对HCoV-OC43易感,HCoV-OC43在其中5种细胞中表现出较强的复制能力,包括BHK-21、293T、Huh7.0、RD等常见贴壁细胞。复制高峰在第5d出现,但HCoV-OC43感染的细胞系在6d内均未观察到典型细胞病变效应。本研究为今后HCoV-OC43的复制规律、致病机制的研究以及筛选广谱抗冠状病毒药物的提供了参考细胞系。
In order to obtain a cell line that can be used for the isolation and research of human coronavirus OC43 (HCoV-OC43), this study used recombinant HCoV-OC43 (rOC43-ns2DelRluc) Nuclear cell lines were susceptible and efficient screening studies, the further use of the parent virus HCoV-OC43-WT were observed cytopathic effect, and the use of quantitative PCR for replication kinetics. The results showed that 18 kinds of cells were susceptible to HCoV-OC43, and HCoV-OC43 showed strong replication ability in 5 kinds of cells, including common adherent cells such as BHK-21,293T, Huh7.0 and RD. The peak of replication appeared on the 5th day, but no HCoV-OC43-infected cell line observed typical cytopathic effect within 6 days. This study provides a reference cell line for future research on the replication of HCoV-OC43, the study of pathogenic mechanism and the screening of broad-spectrum anti-coronavirus drugs.