摘要
目的青岛地区2020年儿童肺炎住院患者人鼻病毒(human rhinovims,HRV)的分离与鉴定,为儿童肺炎的预防提供科学数据。方法收集2020年儿童肺炎住院患者的咽拭子样本,共计98份。应用荧光定量RT-PCR方法进行常见呼吸道病毒筛查,鉴定出HRV阳性的样本接种于H1-HeLa细胞,观察细胞病变。荧光定量RT-PCR鉴定后,扩增HRV-VP4/VP2基因并测序,使用MEGA软件构建系统进化树和同源性分析。结果98例肺炎住院患儿咽拭子样本中HRV阳性11例。接种H1-HeLa细胞,两份样本出现典型的细胞病变。经荧光定量RT-PCR鉴定为HRV病毒。VP4/VP2基因测序结果显示,分离的2株毒株的血清型分别为HRV-A28和HRV-A58。HRV-A28分离株与2011年新加坡参考株、2017年突尼斯参考株和2017年肯尼亚的参考株遗传距离较近,同属一个小分支。HRV-A58分离株与2009澳大利亚、2011年委内瑞拉、2011蒙古和2014年的美国的参考株的遗传距离较近,同属一个小分支。结论本研究首次在青岛地区的儿童肺炎患者咽拭子中分离出HRV-A28和HRV-A58毒株。
Objective To Isolate and identify human rhinovirus(HRV)in hospitalized children with pneumonia in Qingdao in 2020.Methods A total of 98 throat swab samples were collected from hospitalized children with pneumonia in 2020.Common respiratory viruses were screened through RT-qPCR.The HRV positive samples were inoculated into H1-HeLa cells.Viruses with typical cytopathic effect(CPE)were identified by HRV specific RT-PCR.Subsequently,sequences of HRV-VP4/VP2 gene were used to construct phylogenetic trees and analyze homology with sequences of reference strains through MEGA software.Results Among 98 hospitalized children with pneumonia,11 samples were HRV positive in 98 throat swab samples.After the typical CPE appeared in HeLa cells,two strains of HRV were identified by specific RT-PCR.The HRV-A28 and HRV-A58 were confirmed by comparison and analysis of VP4/VP2 sequence.Phylogenetic tree found that the isolated HRV-A28 strain was genetically close to strains of Singapore in 2011,of Tunisia in 2017,and Kenya in 2017.The isolated HRV-A58 strain was genetically close to the strains of Australia in 2009,Venezuela in 2011,Mongolia in 2011,and the United States in 2014.Conclusions The HRV-A28 and HRV-A58 strains were isolated from the throat swabs of children patients with pneumonia in Qingdao.
作者
万以秋
姜法春
宋娟
贾静
王霞
王衍海
梅国勇
蔡茹
韩俊
Wan Yiqiu;Jiang Fachun;Song Juan;Jia Jing;Wang Xia;Wang Yanhai;Mei Guoyong;Cai Ru;Han Jun(School of Medicine,Anhui University of Science and Technology,Huainan 232001,China;Qingdao Center for Disease Control and Prevention,Qingdao 266033,China;State Key Laboratory of Infectious Disease Prevention and Control,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Qingdao Shibei District Center for Disease Control and Prevention,Qingdao 266022,China)
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
2022年第2期219-224,共6页
Chinese Journal of Experimental and Clinical Virology
基金
国家传染病重大专项(2018ZX10102001、2018ZX10711001、2018ZX10734404)
国家病原微生物资源库(NPRC-32)
传染病预防控制国家重点实验室发展基金(2011SKLID104)
安徽理工大学研究生创新基金(2020CX2085)。
关键词
人鼻病毒
病毒分离
系统进化分析
Human rhinovirus
Virus isolation
Phylogenetic analysis
