北京地区1株呼吸道合胞病毒A亚型ON1基因型流行株的分离及其生物学特性研究  

Isolation and characterization of an epidemic ON1 genotype of respiratory syncytial virus subtype A in Beijing

作　　者：单徐畅 任皎[2] 张伟 张钟贤 张菱芳 李佳[4] 楚巧鸿 宋敬东[2] 陈志海 邓瑶[2] 翟德胜[1] 谭文杰[2,4] Shan Xuchang;Ren Jiao;Zhang Wei;Zhang Zhongxian;Zhang Lingfang;Li Jia;Chu Qiaohong;Song Jingdong;Chen Zhihai;Deng Yao;Zhai Desheng;Tan Wenjie(School of Public Health,Xinxiang Medical University,Xinxiang 453003,China;Key Laboratory of Biosafety,National Health Commission of the People′s Republic of China,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Center of Infectious Diseases,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China;Zhejiang Provincial Key Laboratory of Medical Genetics,School of Laboratory Medicine and Life Sciences,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]新乡医学院公共卫生学院,新乡453003 [2]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生物安全重点实验室,北京102206 [3]首都医科大学附属北京地坛医院感染性疾病临床中心,北京100015 [4]温州医科大学检验医学院生命科学学院,浙江省医学遗传学重点实验室,温州325035

出　　处：《中华微生物学和免疫学杂志》2025年第1期10-16,共7页Chinese Journal of Microbiology and Immunology

基　　金：国家重点研发计划(2022YFC2304100)。

摘　　要：目的分离北京地区流行的1株呼吸道合胞病毒(respiratory syncytial virus,RSV),了解该毒株的基因分型和生物学特性。方法从2023年北京地区1例RSV感染患儿鼻咽分泌物标本中分离RSV。qRT-PCR扩增病毒核酸;利用透射电子显微镜、间接免疫荧光试验、空斑形成试验鉴定病毒;根据全基因组测序结果进行基因组系统发育分析;测定病毒滴度,分析复制特点;验证分离毒株用于抗病毒药物体外筛选平台的有效性。结果成功分离1株RSV毒株hRSV/C-Tan/BJ 202301,其感染Hep-2细胞可形成合胞体。电镜下可观察到呈球形、丝状及不规则的病毒颗粒;经免疫荧光检测,病毒感染Hep-2细胞可见绿色荧光,空斑试验结果可见圆斑,形态与Long株相同;基因组序列分析显示其为ON1基因型;与Long株具有相似的细胞生长动力学特性;可应用于抗病毒药物体外筛选。结论本研究从临床样本中成功分离并鉴定1株RSV,该毒株的生物学特性和亲缘关系反映了北京地区流行毒株的特点,为我国RSV疫苗研发及抗病毒药物筛选提供实验材料。ObjectiveTo obtain a prevalent respiratory syncytial virus(RSV)clinical isolate in Beijing and analyze the genotype and biological characteristics of the strain.MethodsA nasopharyngeal secretion specimen was collected from a child with RSV infection in Beijing in 2023 and used for viral isolation.Viral nucleic acid was amplified using qRT-PCR.The isolated virus was identified by transmission electron microscopy,indirect immunofluorescence assay,and plaque formation assay.A phylogenetic analysis was conducted based on the whole-genome sequencing results.Virus titers were determined,and replication characteristics were analyzed.The efficacy of the isolated strain for in vitro screening of antiviral drugs was validated.ResultsA clinical RSV isolate,named hRSV/C-Tan/BJ 202301,was successfully isolated,which could form syncytia in Hep-2 cells.Spherical,filamentous,and irregular virus particles were observed by electron microscopy.Immunofluorescence detection showed green fluorescence in Hep-2 cells,and plaque assay showed round plaques,which were similar to the Long strain in morphology.Genomic sequence analysis showed that it belonged to ON1 genotype.It exhibited similar cell growth kinetics characteristics with the Long strain and could be used for antiviral drug screening in vitro.ConclusionsIn this study,one RSV strain is successfully isolated and identified.The biological characteristics and the phylogenetic relationship of this strain reflect the characteristics of the circulating strains in Beijing,which provides experimental material for RSV vaccine development and antiviral drug screening in China.

关 键 词：呼吸道合胞病毒 病毒分离 鉴定 ON1基因型 

分 类 号：R373[医药卫生—病原生物学]