摘要
目的对安徽蚌埠急性呼吸道感染患者咽拭子样本进行人鼻病毒(human rhinovirus,HRV)的分离与鉴定。方法选择安徽蚌埠哨点医院诊断为急性上呼吸道感染患者的108份咽拭子标本,提取病毒RNA,采用HRV通用引物进行荧光定量PCR检测。将阳性标本进行病毒分离,对HRV病毒株进行HRV VP1基因的扩增与进化树序列分析。结果荧光定量PCR检测HRV呈阳性标本共5份,标本感染H1-Hela细胞3代后只有1份标本出现显著细胞病变效应。此标本经RT-PCR扩增出HRV VP1基因,基因序列分析显示该病毒株与其他代表性HRV A型病毒株的VP1核苷酸同源性分别都在95%以上,命名为TYZQ201901,进化树显示该株病毒与RMH127/2 013株的遗传距离最近,在一个分支上,确认为HRV A型病毒。结论本研究采用H1-Hela细胞顺利从安徽蚌埠急性呼吸道感染患者咽拭子样本分离到1株HRV A型病毒,初步建立了HRV病毒分离技术体系。
Objective To isolate and identify human rhinovirus(HRV)from throat swab samples from patients with acute respiratory infection in Anhui Bengbu.Methods The throat swab specimens from 108 patients with acute upper respiratory tract infection diagnosed in the Anhui Bengbu were selected as samples.RNA was extracted and detected by HRV universal primers;H1-Hela cells were infected with the positive samples for virus isolation,and HRV VP1 gene amplification was performed and gene phylogenetic tree analysis of the successfully isolated HRV strains was done.Results Only 5 samples were positive for HRV by the real-time PCR,and only one sample showed significant cytopathic effects after three passages of H1-Hela cells were infected,and the HRV VP1 gene was amplified by RT-PCR in the sample that was named TYZQ201901.The sequence analysis showed that the VP1 nucleotide homology of the strain with other representative HRV A strains was over 95%.The gene phylogenetic tree analysis showed that the strain had the closest genetic distance to the RMH127/2013 strain,and both were on a branch and was confirmed to be HRV type A virus.Conclusions An HRV type A strain virus was isolated from throat swab samples from patients with acute respiratory infection in Anhui Province by the H1-Hela cells,and the HRV virus separation technology system were initially established.
作者
曹天宇
夏志强
王文军
宋娟
王衍海
韩俊
Cao Tianyu;Xia Zhiqiang;Wang Wenjun;Song Juan;Wang Yanhai;Han Jun(State Key Laboratory of Infectious Disease Prevention and Control,National Institute of Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;School of Public Health,Wannan Medical College,Wuhu 241000,China)
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
2019年第4期358-361,共4页
Chinese Journal of Experimental and Clinical Virology
关键词
急性呼吸道感染
咽拭子
鼻病毒
Acute respiratory infection
Throat swab
Rhinovirus
